News Article | March 1, 2017
It's what's missing in the tumor genome, not what's mutated, that thwarts treatment of metastatic melanoma with immune checkpoint blockade drugs, researchers at The University of Texas MD Anderson Cancer Center report in Science Translational Medicine. Whole exome sequencing of tumor biopsies taken before, during and after treatment of 56 patients showed that outright loss of a variety of tumor-suppressing genes with influence on immune response leads to resistance of treatment with both CTLA4 and PD1 inhibitors. The team's research focuses on why these treatments help 20-30 percent of patients -- with some complete responses that last for years - but don't work for others. Their findings indicate that analyzing loss of blocks of the genome could provide a new predictive indicator. "Is there a trivial or simple (genomic) explanation? There doesn't seem to be one," said co-senior author Andrew Futreal, Ph.D., professor and chair of Genomic Medicine and co-leader of MD Anderson's Moon Shots Program™. "There's no obvious correlation between mutations in cancer genes or other genes and immune response in these patients." "There are, however, pretty strong genomic copy loss correlates of resistance to sequential checkpoint blockade that also pan out for single-agent treatment," Futreal said. Doctoral candidate Whijae Roh, co-lead author, Futreal, and co-senior author Jennifer Wargo, M.D., associate professor of Surgical Oncology and Genomic Medicine, and colleagues analyzed the genomic data for non-mutational effects. "We found a higher burden of copy number loss correlated to response to immune checkpoint blockade and to lower immune scores, a measure of immune activation in the tumor's microenvironment," said Roh, a graduate student in the University of Texas MD Anderson UTHealth Graduate School of Biomedical Sciences. "We also found copy loss has an effect that is independent of mutational load in the tumors." Melanoma tumors with larger volumes of genetic alterations, called mutational load, provide more targets for the immune system to detect and are more susceptible to checkpoint blockade, although that measure is not conclusive alone. "Combining mutational load and copy number loss could improve prediction of patient response," Wargo said. When the team stratified patients in another data set of patients by whether they had high or low copy loss or high or low mutational load, they found that 11 of 26 patients with high mutational load and low copy loss had a clinical benefit, while only 4 or 26 with low mutational load and high copy loss benefited from treatment. In the trial, patients were treated first with the immune checkpoint inhibitor ipilimumab, which blocks a brake called CTLA4 on T cells, the immune system's specialized warriors, freeing them to attack. Patients whose melanoma did not react then went on to anti-PD1 treatment (nivolumab), which blocks a second checkpoint on T cells. Biopsies were taken, when feasible, before, during and after treatment for molecular analysis to understand response and resistance. To better understand the mechanisms at work, the team analyzed tumor genomes for recurrent copy loss among 9 tumor biopsies from patients who did not respond to either drug and had high burden of copy number loss. They found repeated loss of blocks of chromosomes 6, 10 and 11, which harbor 13 known tumor-suppressing genes. Analysis of a second cohort of patients confirmed the findings, with no recurrent tumor-suppressor loss found among any of the patients who had a clinical benefit or long-term survival after treatment. Ipilimumab sometimes wins when it fails The researchers also found a hint that treatment with ipilimumab, even if it fails, might prime the patient's immune system for successful anti-PD1 treatment. The team analyzed the genetic variability of a region of the T cell receptors, a feature of T cells that allows them to identify, attack and remember an antigen target found on an abnormal cell or an invading microbe. They looked for evidence of T cell "clonality," an indicator of active T cell response. Among eight patients with longitudinal samples taken before treatment with both checkpoint types, all three who responded to anti-PD1 therapy had shown signs of T cell activation after anti-CTLA treatment. Only one of the five non-responders had similar indicators of T cell clonality. "That's evidence that anti-CTLA4 in some cases primes T cells for the next step, anti-PD1 immunotherapy. It's well known that if you don't have T cells in the tumor, anti-PD1 won't do anything, it doesn't bring T cells into the tumor," Futreal says. Overall, they found that T cell clonality predicts response to PD1 blockade but not to CTLA-4 blockade. "Developing an assay to predict response will take an integrated analysis, thinking about genomic signatures and pathways, to understand the patient when you start therapy and what happens as they begin to receive therapy," Wargo said. "Changes from pretreatment to on-therapy activity will be important as well." The Science Translational Medicine paper is the third set of findings either published or presented at scientific meetings by the team, which is led by Futreal and Wargo, who also is co-leader of the Melanoma Moon Shot™. Immune-monitoring analysis showed that presence of immune infiltrates in a tumor after anti-PD1 treatment begins is a strong predictor of success. They also presented evidence that the diversity and composition of a patient's gut bacteria also affects response to anti-PD1 therapy. The serial biopsy approach is a hallmark of the Adaptive Patient-Oriented Longitudinal Learning and Optimization™ (APOLLO) platform of the Moon Shots Program™, co-led by Futreal that systematically gathers samples and data to understand tumor response and resistance to treatment over time. The Moon Shots Program™ is designed to reduce cancer deaths by accelerating development of therapies, prevention and early detection from scientific discoveries. Futreal holds the The Robert A. Welch Distinguished University Chair in Chemistry at MD Anderson. Co-authors with Roh, Futreal and Wargo are co-first authors Pei-Ling Chen, M.D., of Genomic Medicine and Pathology, and Alexandre Reuben, Ph.D., of Surgical Oncology; also Christine Spencer, Feng Wang, Ph.D., Zachary Cooper, Ph.D., Curtis Gumbs, Latasha Little, Qing Chang, Wei-Shen Chen, M.D., and Jason Roszik, Ph.D., of Genomic Medicine; Michael Tetzlaff, Ph.D., M.D., and Victor Prieto, M.D., Ph.D., of Pathology; Peter Prieto, M.D., Vancheswaran Gopalakrishnan, Jacob L. Austin-Breneman, Hong Jiang, Ph.D., and Jeffrey Gershenwald, M.D., of Surgical Oncology; John Miller, Ph.D., Oncology Research for Biologics and Immunotherapy Translation (ORBIT); Sangeetha Reddy, M.D., Division of Cancer Medicine; Khalida Wani, Ph.D., Mariana Petaccia De Macedo, M.D., Ph.D., Eveline Chen, and Alexander Lazar, M.D., Ph.D., of Translational Molecular Pathology; Michael Davies, M.D., Ph.D., Hussein Tawbi, M.D., Ph.D., Patrick Hwu, M.D., Wen-Jen Hwu, M.D., Ph.D., Adi Diab, M.D., Isabella Glitza, M.D., Ph.D., Sapna Patel, M.D., Scott Woodman, M.D., Ph.D., and Rodabe Amaria, M.D., of Melanoma Medical Oncology; Jianhua Hu, Ph.D., of Biostatistics; Padmanee Sharma, M.D., Ph.D., and James Allison, Ph.D., of Immunology; Lynda Chin, M.D., University of Texas System; and Jianhua Zhang Ph.D., of the Institute for Applied Cancer Science. Wargo, Sharma and Allison are all members of the Parker Institute for Cancer Immunotherapy. The research was funded by MD Anderson's Melanoma Moon Shot™, the Melanoma Research Alliance Team Science Award, the John G. and Marie Stella Kenedy Memorial Foundation, the University of Texas System STARS program; the Cancer Prevention and Research Institute of Texas; the American Society of Clinical Oncology; Conquer Cancer Foundation; the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation; and grants from the National Cancer Institute of the National Institutes of Health (U54CA163125, 1K08CA160692-01A1, T32CA009599, NIH T32 CA009666, R01 CA187076-02) and MD Anderson's Institutional Tissue Bank (2P30CA016672) Spencer and Gopalakrishnan are graduate students in The University of Texas Health Science Center at Houston School of Public Health.
News Article | December 15, 2016
This rapidly fatal brain cancer has seen only two improvements in therapy in 30 years BIRMINGHAM, Ala. - In a paper published today in Cancer Research, researchers: 1) identify a biomarker enzyme associated with aggressive glioma brain tumors, 2) reveal the regulatory mechanism for that enzyme, and 3) demonstrate potent efficacy, using a mouse model of glioma, for a small molecule inhibitor they have developed. The inhibitor, GA11, retains a core structure that resembles natural inhibitors of the biomarker enzyme; but the inhibitor has been modified to help it pass through the blood-brain barrier. "In principle, both these features make GA11 an attractive drug candidate to target glioma stem-like cells in glioblastoma multiforme tumors," said Ichiro Nakano, M.D., Ph.D., and colleagues in the paper. Nakano, a professor of neurosurgery and academic neurosurgeon at the University of Alabama at Birmingham, and Vito Coviello and Concettina La Motta, University of Pisa, Italy, are doing further preclinical evaluation of the GA11 and its analogs. Glioblastoma multiforme, or GBM, is a formidable cancer foe. Only two therapeutic improvements have appeared in the past 30 years, increasing the average survival of patients from five months to 15 or 16 months, Nakano says. A GBM tumor is a mix of different cells that respond differently to therapies. Small numbers of glioma stem-like cells, or GSCs, drive the tumorigenicity of GBM and thus are prime targets for possible treatments. One GSC subtype called the mesenchymal GSC is more malignant and the most therapy-resistant, so Nakano and fellow researchers reasoned that identifying the regulatory molecules active in mesenchymal GSCs might lead to novel and effective therapeutics. Nakano and colleagues found that one form of the enzyme aldehyde dehydrogenase -- ALDH1A3 -- is a specific marker for mesenchymal GSCs, and his group is the first to show that, among the heterogeneous mix of cells in a GBM tumor, cells with high levels of ALDH1A3 expression were more tumorigenic in vivo than are cells that are low in ALDH1A3. The researchers also found that the FOXD1 transcription factor regulates the production of ALDH1A3 in mesenchymal GSCs. In clinical samples of high-grade gliomas from patients, the expression levels of both FOXD1 and ALDH1A3 were inversely correlated with disease progression -- gliomas with high levels were more rapidly fatal than were gliomas with low levels. Astonishingly, the same mechanism that drives the mesenchymal GSC tumorigenicity in humans acts in an evolutionarily distant organism, the fruit fly. Knocking down the expression of either the fruit fly version of the FOXD1 gene or the fruit fly version of ALDH1A3 blocks the formation of brain tumors in a brain cancer model of the fruit fly species Drosophila melanogaster, the researchers found. Thus, this signaling has been highly conserved in evolution. The FOXD1 transcription factor is normally active during development from a fertilized egg and embryo to a fetus, and it is silent after birth. The role of FOXD1 in GBM, Nakano and colleagues say, suggests that the mesenchymal GSCs have hijacked the molecular mechanism of normal embryonic development to promote tumor growth. In preclinical testing, GA11 was validated several ways. The researchers showed that it inhibited ALDH in yeast, reduced ALDH1 activity in cell-culture spheres of mesenchymal GSCs, inhibited proliferation of glioma spheres in cell culture, and inhibited xenograft growth of GBM in mouse brains. "In conclusion," Nakano and fellow researchers wrote, "the FOXD1-ALDH1A3 axis is critical for tumor initiation in mesenchymal GSCs, therefore providing possible new molecular targets for the treatment of GBM and other ALDH1-activated cancers." Nakano says his study of the role of GSCs in GBM is just one approach to treat glioma tumors. Other labs are pursuing immunotherapy, the use of check-point inhibitors, vaccination and efforts to increase sensitivity to radiotherapy. It will take combined therapies to treat glioblastoma, Nakano says. "We don't believe that one therapy will be effective." Nakano expects to launch a new clinical trial for glioblastoma in 2017, in conjunction with Burt Nabors, M.D., professor of neurology at UAB. Nakano says UAB will be the only site in the Deep South for this unique trial aimed at a molecular target in glioma stem cells, a target that is different from the ones described in the Cancer Research paper. The referral contact to Nakano's service will be Lydia P. Harrell. The Nakano lab is also working on brain metastases, tumors that spread into the brain from other parts of the body. Similar to high-grade gliomas, which originate in the brain, these metastatic brain tumors are lethal, and there are very few therapeutic options. Nakano believes the core stem cell genes and signaling pathways are shared between gliomas and brain metastases. "If so," he said, "the molecular targets identified for gliomas are most likely essential in brain metastases. Studies are underway, and similar to the glioma therapy development, I am working to develop clinical trials for brain metastasis, together with medical oncologists Mansoor Saleh, M.D., Andres Forero, M.D., and others at UAB." Besides Nakano, Coviello and La Motta, authors of the Cancer Research paper, "FOXD1-ALDH1A3 signaling is a determinant for the self-renewal and tumorigenicity of mesenchymal glioma stem cells," are Peng Cheng, Jia Wang, Zhuo Zhang, Sung-Hak Kim, Marat S. Pavlyukov and Mutsuko Minata, UAB Department of Neurosurgery and Comprehensive Cancer Center; Indrayani Waghmare and Madhuri Kango-Singh, Department of Biology, University of Dayton, Ohio; Stefania Sartini, Department of Pharmacy, University of Pisa, Italy; Ahmed Mohyeldin, Department of Neurological Surgery and the James Comprehensive Cancer Center, The Ohio State University; Claudia L.L. Valentim, Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic; Rishi Raj Chhipa and Biplab Dasgupta, Department of Oncology, Cincinnati Children's Hospital Medical Center; and Krishna P.L. Bhat, Department of Translational Molecular Pathology, The University of Texas, M.D. Anderson Cancer Center, Houston.
News Article | February 28, 2017
Adding two blood-borne proteins associated with cancer cell migration increases the predictive ability of the current biomarker for pancreatic cancer to detect early stage disease, a research team from The University of Texas MD Anderson Cancer Center reports in the Journal of the National Cancer Institute. The trio of biomarkers, tested in three separate cohorts, including two blinded validation studies, improved the detection of patients with early stage disease compared to healthy or benign disease controls. "Adding these two biomarkers provided statistically significant improvement for all early stage cancer versus healthy controls as well as other subcohorts when used with the current gold standard biomarker, CA 19-9," said Ann Killary, Ph.D., professor of Translational Molecular Pathology. While CA 19-9 is the only biomarker approved for use by the U.S. Food and Drug Administration, and only for monitoring treatment for the disease, according to first author Seetharaman Balasenthil, Ph.D., instructor in Translational Molecular Pathology. The marker of antigens produced by pancreatic cancer has a low positive predictive value for identifying early stage disease. At early stages, pancreatic cancer can be successfully removed with surgery, but 80 percent of patients are diagnosed with either locally advanced disease (stage III) or cancer that has spread to other organs (stage IV), when surgery is no longer a curative option. Pancreatic cancer is the fourth-leading cause of deaths from cancer in the United States, with an estimated 53,070 new cases diagnosed in 2016 and 41,780 deaths, according to the National Cancer Institute. "Our goal is to identify more patients at those earlier, resectable stages, when treatment could lead to a five year survival rate of 30 percent or more, depending on stage," Killary said. Only about 7 percent of patients survive for five years following diagnosis of the disease. Additional studies in larger cohorts will be needed to validate these findings, and more biomarkers will be needed to get the completely accurate set needed to screen the general population with the long term goal of identifying precursor lesions before they become malignant. Killary and colleagues earlier identified a cluster of genes involved in cancer migration and then analyzed the proteins produced by those genes. Two, known as TFPI and tenascin C, emerged as the strongest biomarker candidates. The team first compared their predictive ability by comparing their presence in healthy volunteers and those with primarily stage IV pancreatic cancer. In the current study, deploying TFPI and an isoform of tenascin C, TNC-FN IIIC, with CA 19-9 improved performance discriminating stage I and II disease from healthy or benign disease controls. The principal measure of efficacy is called Area Under the Curve (AUC), a measure of how well biomarkers identify true cases of disease (sensitivity) while avoiding false positives (specificity). A perfect AUC would have a score of 1.0. Any biomarker test for the general public will need to be close to a perfect 1.0, Killary noted, given the infrequency of pancreatic cancer in the population at large. In the group's analysis of three cohorts of samples, AUC scores for the combination consistently outscored CA-19-9 alone. For the first cohort of samples, comparing stage I/IIA and healthy controls yielded an AUC of 0.72 for CA-19-9 alone compared to 0.84 for the combination. For stage IIB vs. controls, the combination raised CA-19-9's performance from an AUC of 0.87 to 0.98. The second blinded cohort also showed improvements in AUC over CA19-9. A third, larger blinded validation study of 252 samples of cases and controls from the Early Detection Research Network of the National Cancer Institute further confirmed those findings and also permitted the team to measure the trio's ability to detect disease in patients without a history of pancreatitis or diabetes. The AUC for those patients was 0.87 for Stage I/IIA, 0.93 for Stage IIB and 0.89 for all early stages of the disease using the combination, pointing to a potential ability to detect disease in those who lack either of those known risk-raising conditions. Killary said the team is working to use the biomarkers in MD Anderson's high-risk clinic, established through MD Anderson's Pancreatic Cancer Moon Shot™, part of the institution's Moon Shots Program™ to accelerate the development of life-saving advances based on scientific discovery. The clinic monitors people who are already at high risk of pancreatic cancer due to family history or having known risk-raising genetic mutations. "In this population, our biomarker panel might prove very useful in early detection," Killary said. Co-author Subrata Sen, Ph.D., also a professor of Molecular Translational Pathology, notes, "These findings are a significant advance considering that there's nothing else available now to detect early stage pancreatic cancer that has gone through blinded validations in multiple patient cohorts." Co-authors with Killary, Balasenthil and Sen are Nanyue Chen, M.D., Ph.D., of Translational Molecular Pathology; Suyu Liu, Ph.D., and J. Jack Lee, Ph.D., of Biostatistics; Jinyun Chen, M.D., Pharm.D., and Marsha Frazier, Ph.D. of Epidemiology; Ying Huang, Ph.D., and Tracey Marsh, of Fred Hutchinson Cancer Research Center, Seattle; Sanford Stass, M.D., and Debra KuKuruga , Ph.D., of the University of Maryland Medical Center, Baltimore; Randall Brand, M.D., University of Pittsburgh Medical Center; and Sudhir Srivastava, M.D., of the National Cancer Institute. This research was funded by Early Detection Research Network grants (U01 CA111302 and U24 CA115091-10) and by MD Anderson's Moon Shots Program.
News Article | February 20, 2017
Jatkoimme jakelijaverkostomme laajentamista uusilla sopimuksilla ja sopimusten uudelleenjärjestelyillä. Vuoden 2016 aikana solmimme seuraavat sopimukset Biohitin diagnostiikkatuotteiden jakelusta: OnSite Diagnostic Lab India Pvt Ltd myy diagnostisia testejämme Intiassa ja SPD Scientific Pte Ltd. Thaimaassa. Filippiineillä diagnostisia testejä jakelee jatkossa UC Biosciencies Inc. GastroPanel®-testin jakelijaksi Iranissa nimettiin Pooyandegan Pezeshki Pardis ja Delta Biologicals S.r.l. Kaakkois-Italiassa. Biohitin kalprotektiinitestin jakelijaksi nimitettiin Kiinassa Tianjin Jingsheng Biological Technology Development Co. Ltd. Jatkoimme venäläisen kumppanimme Melon OOO:n kanssa diagnostiikkatuotteidemme yksinmyyntisopimusta seuraavalle sopimuskaudelle. Sovimme myös Beijing HuayiHuilang Medical Instrument Co., Ltd:n kanssa keliakiapikatestin ja GA-mapTM-dysbioositestin jakelusta Kiinassa sekä Glomedics:n kanssa laktoosi-intoleranssipikatestin jakelusta Etelä-Koreassa. Sri Lankassa helikobakteeripikatestin jakelijaksi nimitettiin Easemed Global Ltd. Lisäksi iranilaisen Arad Tajhiz Azma:n kanssa tehtiin sopimus kalprotektiini-testin yksinoikeudellisesta jakelusta Iranissa. Ruotsissa diagnostisia testejä jakelee XboXLab AB. Biohit Oyj:n kiinalaisen yhteisyrityksen Biohit HealthCare (Hefei) Co. Ltd:n tuotantolaitos läpäisi viranomaisten asettamat vaatimukset testeille ja sai toimiluvan, joka mahdollistaa tuotteiden tuotannon ja myynnin. Biohit HealthCare (Hefei) Co. Ltd valmistaa Biohit Oyj:n kehittämiä GastroPanel®-tuotteita Kiinan markkinoille. Tappiollisista tilikausista huolimatta yhtiö on onnistunut pitämään käyttöpääomansa hyvällä tasolla ja yhtiön johto uskoo, että käyttöpääoma riittää kattamaan seuraavien 12 kuukauden toiminnan, eikä yhtiö ole riippuvainen ulkopuolisesta rahoituksesta liiketoiminnan jatkuvuuden takaamiseksi. Yhtiö odottaa lisäksi saavansa 2.1.2017 tiedotetusta yhteisyrityksen omistusjärjestelystä noin 1,8 milj. euroa käteistä ensimmäisen vuosipuoliskon aikana, jonka odotetaan osaltaan vahvistavan yhtiön käyttöpääomarakennetta. Omistusjärjestelyn toteutuminen edellyttää viranomaishyväksyntää. Yhtiön johdon arvio on, että yhtiön kyky jatkaa toimintaansa on hyvä, eikä sellaisia tapahtumia tai olosuhteita ole tiedossa, jotka yksin tai yhdessä saattavat antaa merkittävää aihetta epäillä yhteisön kykyä jatkaa toimintaansa. Loppuvuonna 2015 päätökseen saadun seniori-tutkimuksen tulokset julkaistiin vuoden 2016 aikana Journal of Aging Research and Clinical Practice-tiedelehdessä. Tässä tutkimuksessa vertailtiin yli 200 vanhusten palvelutaloissa asuvaa, keski-iältään yli 80-vuotiasta henkilöä Suomessa ja Virossa (Tampere, Tartto) selvittämällä GastroPanel®- ja B12-vitamiinitestillä mm. atrofisen gastriitin, helikobakteeri-infektion ja B12-vitamiinipuutoksen yleisyys.Tutkimuksessa todettiin huomattavat erot näiden kahden maan vanhusväestön yleisessä terveydentilassa, sekä erityisesti helikobakteeri-infektion ja B12-vitamiinipuutoksen esiintyvyydessä, toteamisessa sekä niiden hoitokäytännöissä. GastroPanel®-tutkimuksella löydettävä atrofinen gastriitti on yleinen B12-vitamiinin imeytymishäiriön syy, jonka varhaisen toteamisen merkitys korostuu myös tässä tutkimuksessa. Biohit Oyj:n kliininen tutkimusosasto on käynyt läpi vuoden 2016 aikana kaikki tähän mennessä kertyneet tieteelliset julkaisut, joissa on tutkittu GastroPanel®-testin käyttöä atrofisen gastriitin diagnostiikassa ympäri maailmaa. Kaikki tietyt valintakriteerit täyttävät tutkimukset sisällytettiin ns. meta-analyysiin. Meta-analyysi on tilastollinen analyysimenetelmä, jonka avulla voidaan laskea mm. se, mikä on GastroPanel®-testin herkkyys ja tarkkuus atrofisen gastriitin diagnostiikassa, huomioiden kaikki julkaistut tieteelliset raportit. Valintakriteerit täytti kaikkiaan 27 tutkimusta, joissa on tutkittu lähes 9000 potilasta eri maissa ja maanosissa. Huolimatta yksittäisten tutkimusten välisestä huomattavastakin vaihtelusta, meta-analyysin antamat kokonaistulokset ovat rohkaisevia ja selvästi tukevat Biohitin omissa ja sen partnereiden tekemissä tutkimuksissa saatuja positiivisia kokemuksia testistä. Erityisen tärkeä on GastroPanel®-testin atrofista gastriittia koskeva korkea tarkkuus, mikä käytännössä tarkoittaa, että testin ollessa täysin normaali, on todennäköisyys atrofiselle gastriitille tai helikobakteeri-infektiolle mahalaukussa häviävän pieni. Paitsi tätä Anticancer Research-lehdessä julkaistua meta-analyysiä, vuoden 2016 aikana on samassa lehdessä raportoitu myös kaksi muuta Biohitin ja/tai sen yhteistyökumppanien tekemää tutkimusta, joiden tulokset ovat merkittäviä. Näistä toinen on Pietarissa toteutettu oireettomien henkilöiden GastroPanel®-seulonta, ja toinen on tapaus-verrokkiasetelmassa toteutettu kymmenen vuoden seurantatutkimus, jossa selvitettiin GastroPanel®-testin arvoa pitkän aikavälin mahasyöpäriskin ennustajana. GastroPanel®-testin käyttökelpoisuus oireettoman väestön seulontamenetelmänä oli erinomainen ja testi oli herkkä ja tarkka löytämään mm. atrofisen gastriitin (www.biohit.fi/lisatietoja ). Tapaus-verrokki-tutkimuksessa GastroPanel®-merkkiaineiden (PGI ja PGII) poikkeavan matalat arvot olivat itsenäinen kohonneen mahasyöpäriskin ennustaja jopa kymmenen vuoden seurantajakson aikana. Monimuuttujamallissa kaikkein vahvin ennustaja oli matala PGI/PGII suhde, joka GastroPanel®-testin tulkinnassa on yksi limakalvoatrofian indikaattori. Vuoden 2016 aikana tietoisuutta asetaldehydin haitallisuudesta edistettiin useissa eri yhteyksissä. Vuoden 2016 Lääkäripäivien yhteydessä professori Mikko Salaspuro piti esityksen asetaldehydistä maailman yleisimpänä karsinogeeninä. Esitys herätti vilkkaan julkisen keskustelun aiheesta. Samasta aiheesta professori Salaspuro piti esitelmän myös kahdeksannessa Charles Lieber satelliitti symposiumissa New Orleansissa 25 kesäkuuta 2016. Esitelmän nimi oli ”Acetaldehyde a neglected human carcinogen” ja sen lyhennelmä on julkaistu kansainvälisessä julkaisusarjassa ”Experimental and Molecular Pathology 102: 2017”. Nykytiedon valossa alkoholijuomien pakkauksissa tulisi olla merkintä niiden mahdollisesti sisältämästä asetaldehydistä, jonka maailman terveysjärjestö (WHO) luokittelee ihmiselle syöpävaaralliseksi yhdisteeksi. Koska tieteellistä näyttöä ei ole olemassa siitä, että muiden elintarvikkeiden sisältämä asetaldehydi ja etanoli olisivat vähemmän syöpävaarallisia kuin alkoholijuomissa oleva asetaldehydi, sama merkintävaatimus tulisi koskea myös niitä. Monien alkoholijuomien ja elintarvikkeiden asetaldehydipitoisuudet ylittävät 5mg/l, joka on kosmeettiselle tuotteelle suositellun asetaldehydipitoisuuden yläraja. Lisäksi EU:n asettama tieteellinen komitea on esittänyt vuonna 2012, että suuvesissä asetaldehydiä ei saisi olla lainkaan. Kiinan liiketoiminta on vuonna 2017 Biohitin fokuksessa. Käynnissä olevien seulontatutkimusten lisäksi Biohitin yhteisyritys Biohit HealthCare (Hefei) Co. Ltd on onnistunut rakentamaan kattavan jakeluverkoston ja asiakaspohjan sekä kaupallistamaan GastroPanel® -tuotteen laajalti hyödyntäen työtä, joka on tehty hintahyväksyntöjen saamiseksi Kiinan maakuntatasolla aikaisemmin. Myyntikehitys on ollut hyvää ja nousujohteista erityisesti toisella vuosipuoliskolla. Yhteisyritys on hyvin asemoitu jatkamaan tätä kehitystä myös vuonna 2017. Biohit odottaa saavansa päätökseen yhteisyrityksen osakepääoman alentamisprosessin ensimmäisen vuosineljänneksen aikana. Transaktion seurauksena Biohit Oyj arvioi vuoden 2017 ei-vertailukelpoisen liiketuloksen kääntyvän selvästi voitolliseksi. Yhteisyrityksen osakkuudesta luopuminen on osa tulevaisuuden GastroPanel® -strategiaamme Kiinassa. Tuotantolaitoksen perustaminen, vaadittavien lupien saaminen ja uuden tuotteen kaupallistaminen edellytti riskin jakamista yhteisyrityksen muodossa, mutta kokonaan kiinalaisomisteisen yhtiön on helpompi päästä kansallisiin terveydenhoito-ohjelmiin ja kasvun rahoitukseen, kuin osittain ulkomaalaisomisteisen yhtiön. Ensimmäinen meta-analyysi GastroPanel®-testillä tehdyistä tieteellisistä tutkimuksista valmistui. GastroPanel® on ainutlaatuinen, runsasti tietoa antava testi ylävatsavaivoista kärsivien tutkimiseen ja helikobakteeri-gastriitin (-infektion) ja siitä kehittyneen mahalaukun limakalvon surkastuman eli atrofisen gastriitin ja siitä johtuvien mahasyöpä- ym. riskien toteamiseen. Vain GastroPanelilla® tai gastroskopia- ja koepalatutkimuksella voidaan löytää atrofinen gastriitti, joka on useimmiten oireeton ja jota siihen liittyvine riskeineen 13C ureahengitystesti, ulosteen antigeenitesti ja pelkät vasta-ainetestit eivät löydä. Mahdollisesti juuri tästä syystä atrofisesta gastriitista kehittynyt mahasyöpä todetaan edelleenkin useimmiten vasta myöhäisvaiheessa, jolloin hoitoennuste on erittäin huono (www.biohit.fi/lisatietoja ). Biohit Oyj:n kiinalaisen yhteisyrityksen Biohit HealthCare (Hefei) Co. Ltd:n tuotantolaitos läpäisi viranomaisten asettamat vaatimukset testeille ja sai toimiluvan, joka mahdollistaa tuotteiden tuotannon ja myynnin. Biohit HealthCare (Hefei) Co. Ltd aloitti Biohit Oyj:n kehittämien maailmassa ainulaatuisten GastroPanel®-tuotteen valmistamisen Kiinan markkinoille, ja ensimmäiset toimitukset menivät käynnissä olevan mahasyövän riskin seulontatutkimuksen tarpeisiin. Biohit Oyj on globaaleilla markkinoilla toimiva suomalainen bioteknologiayritys. Biohitin missio on "Innovating for Health” - tuotamme innovatiivisia tuotteita ja palveluita edistämään tutkimusta ja varhaista diagnostiikkaa. Biohitin pääkonttori on Helsingissä, ja tytäryhtiöt sijaitsevat Italiassa ja Iso-Britanniassa. Biohitin B-sarjan osakkeet (BIOBV) noteerataan Nasdaq Helsingin Small cap/Terveydenhuolto-ryhmässä. www.biohit.fi
News Article | November 9, 2016
SOUTH SAN FRANCISCO, Calif., Nov. 09, 2016 (GLOBE NEWSWIRE) -- Fluidigm Corporation (NASDAQ:FLDM) and GenomOncology today announced a distribution agreement to co-market the GO Clinical Workbench™ for molecular interpretation of somatic variants identified using Fluidigm systems. An elegant solution for clinical research laboratories seeking accuracy, flexibility, and scalability, this combined offering will enable laboratories to achieve greater workflow efficiencies and productivity from NGS library preparation, variant annotation and reporting. Fluidigm currently provides highly-scalable workflows for NGS library preparation that offer significant cost savings and workflow improvements over the largely manual, low-throughput library preparation workflows available today. Utilizing the Juno™ Targeted DNA Sequencing Library Preparation System, a wide range of panels can easily be produced, from targeted multiplexed panels covering a few user‑defined genes to more comprehensive panels of 4,800 or more amplicons from hundreds of genes. With the additional flexibility to add new markers over time, laboratories can also be well-positioned to meet the demands of NGS library preparation today and well into the future. Fluidigm plans to further expand this offering by developing NGS somatic mutation panels together with optimized automated library preparation protocols for use with the Juno system. GenomOncology and Fluidigm plan to co-market the Juno targeted NGS library preparation workflow for use with the GO Clinical Workbench, a proven knowledge-enabled software solution that provides decision support to enable precision oncology research programs. The GO Clinical Workbench is built upon a foundation of expert knowledge curated by My Cancer Genome, which provides information related to cancer genes and variants, targeted therapies, prognostic implications, clinical trials, and cell-signaling pathways. This knowledge base, augmented with information from FDA, National Comprehensive Cancer Network (NCCN), American Society of Clinical Oncology (ASCO), and other annotation sources, is readily available via the GO Clinical Workbench. In support of this partnership, Fluidigm and GenomOncology plan to co-present in the Fluidigm Corporate Workshop at the American Molecular Pathology (AMP) annual meeting on November 9 in Charlotte, North Carolina. “We are pleased to partner with Fluidigm to provide an integrated offering that will enable more laboratories to meet the increasing demands of molecular tumor screening and reporting,” said Manuel Glynias, CEO of GenomOncology. “Fluidigm is well-recognized for delivering trusted performance while providing significant improvements in laboratory productivity, a goal that we both share.” “GenomOncology is leading the way in developing easy-to-use tools to streamline somatic mutation profiling and variant reporting,” said Chris Linthwaite, President and CEO of Fluidigm. “We look forward to providing these industry-leading tools to our customers and further expanding this combined offering with NGS oncology panels for translational and clinical research. Together with our customers, we look forward to seeing the benefits these solutions will bring to the cancer community.” Use of Forward-Looking Statements This press release contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995, including, among others, statements relating to features and benefits of a combined product offering and benefits from, and activities under, a co-marketing agreement. Forward-looking statements are subject to numerous risks and uncertainties that could cause actual results to differ materially from currently anticipated results, including, but not limited to, risks relating to the future financial performance of Fluidigm product lines and challenges inherent in developing, manufacturing, launching, marketing, and selling new products, including reliance on third parties for co-marketing and collaboration and competition. Information on these and additional risks, uncertainties, and other information affecting Fluidigm business and operating results are contained in Fluidigm’s Quarterly Report on Form 10-Q for the quarter ended September 30, 2016, and in its other filings with the Securities and Exchange Commission. These forward-looking statements speak only as of the date hereof. Fluidigm disclaims any obligation to update these forward-looking statements except as may be required by law. About Fluidigm Fluidigm (NASDAQ:FLDM) develops, manufactures, and markets life science analytical and preparatory systems for growth markets such as single-cell biology and production genomics. We sell to leading academic institutions, clinical laboratories, and pharmaceutical, biotechnology, and agricultural biotechnology companies worldwide. Our systems are based on proprietary microfluidics and multiparameter mass cytometry technology and are designed to significantly simplify experimental workflow, increase throughput, and reduce costs while providing excellent data quality. Fluidigm products are provided for Research Use Only. Not for use in diagnostic procedures. We use our website (www.fluidigm.com), corporate Twitter account (@fluidigm), Facebook page (https://www.facebook.com/Fluidigm), and LinkedIn page (https://www.linkedin.com/company/fluidigm-corporation) as channels of distribution of information about our products, our planned financial and other announcements, our attendance at upcoming investor and industry conferences, and other matters. Such information may be deemed material information, and we may use these channels to comply with our disclosure obligations under Regulation FD. Therefore, investors should monitor our website and our social media accounts in addition to following our press releases, SEC filings, public conference calls, and webcasts. Fluidigm, the Fluidigm logo, and Juno are trademarks or registered trademarks of Fluidigm Corporation. About GenomOncology GenomOncology is a health care technology company enabling precision medicine by transforming genomic data into actionable information. Learn more at www.genomoncology.com.
News Article | February 20, 2017
Biohit Oyj Financial Statement Release, 20 February 2017 at 9:30 am local time (EET) ‘In the second half of 2016 our net sales grew by 72.8% compared to the comparison period 7-12/2015. During the entire year our net sales grew by 35.4% compared to 1-12/2015. In terms of operational development, Biohit Oyj has focused on supporting the business activities of our joint venture Biohit HealthCare (Hefei) Co. Ltd, expanding the international distributor network and supporting partners, for example in product registrations. Especially our operations in China developed in a favorable manner in 2016 and our joint venture exceeded EUR 5 million in net sales with GastroPanel® (EUR 0.2 million in 2015) and made a positive operational result (EUR -1.1 million 2015). Our most important products are GastroPanel®, Acetium and diagnostic quick tests. Our main market areas are Asia and Europe. The smoking intervention study, which started in 2016, burdened the financial year’s costs in 2016 but other comparable non-IFRS costs were at a similar level than 2015. For 2017 the costs are not expected to grow, as the cost burden associated with clinical studies focused on the year 2016. In addition the net cash flow from operating activities improved by EUR 0.6 million compared to 2015.’ We expanded our distributor network and advanced product registrations We continued expanding our distributor network with new agreements and by restructuring agreements. During 2016 we signed following distribution agreements of Biohit`s diagnostic products: OnSite Diagnostic Lab India Pvt Ltd sells our diagnostic tests in India and SPD Scientific Pte Ltd. in Thailand. In the Philippines UC Biosciencies Inc. will distribute diagnostic tests. Pooyandegan Pezeshki Pardis was appointed as the GastroPanel® distributor in Iran and Delta Biologicals S.r.I in southeast Italy. Tianjin Jingsheng Biological Technology Development Co. Ltd was appointed as distributor for Biohit´s calprotectin-test in China. We renewed the agreement with our Russian partner Melon OOO for the exclusive right to distribute our diagnostic products to a further contract period. In addition, we made a celiac disease quick test and GA-mapTM dysbiosis test distribution agreements with Beijing HuayiHuilang Medical Instrument Co.,Ltd in China and Glomedics distributes Lactose Intolerance Quick Tests in South Korea. In Sri Lanka Easemed Global Ltd was appointed as distributor for Helicobacter Pylori Quick Test. Furthermore, an agreement with exclusive right to distribute calprotectin-test in Iran was made with Iranian Arad Tajhiz Azma. In Sweden diagnostic tests are distributed through XboXLab AB. During the reporting period Ericon S.r.l. in Moldova and Inversiones Naturalia S.A in Panama got exclusive rights to sell Acetium. The acetaldehyde binding L-cysteine capsule, which was registered in Mexico as a food supplement and under a product name Etium, was granted an import license in Mexico in May 2016. The first batch was delivered to Mexico in early summer. During the first half of 2016 several diagnostic test registrations were concluded in Costa Rica. In the end of 2016 Indian authorities granted import license for Biohit´s several diagnostic tests. Additionally, during the second half of 2016 registrations in Thailand for several diagnostic tests, a re-registration of GastroPanel® and H. Pylori quick test in Kazakhstan and registration of ColonView, UFT300 and Quick test reader in Israel were concluded. During 2016 a price approval decision on the three GastroPanel® tests (pepsinogen l, pepsinogen II, gastrin-17) was issued in four Chinese provinces. The price approval decision has already been issued in 20 provinces. Price approval is a pre-requisite for reimbursability of GastroPanel® and start of sales. In general, the duration of the product registration process is different in each market area. For this reason, it is not possible to accurately assess the time it takes for the authorities to handle registrations in different countries and for product sales to begin. We bought a share of Norwegian Genetic Analysis AS with a directed share issue Biohit Oyj and Genetic Analysis AS signed a share exchange agreement through which Biohit Oyj acquired ownership of 18% of shares in the company. In addition to this the companies signed a distribution agreement giving Biohit Oyj a right to sell Genetic Analysis AS’s Dysbiosis Test under the Biohit brand and exclusively in Finland and in China. In the future, Genetic Analysis will also operate as a distributor to Biohit Oyj’s products and services in Norway. Biohit Oyj’s Chinese joint venture Biohit HealthCare (Hefei) Co. Ltd production facility has passed official test requirements and has been granted a license enabling production and sales of products. Biohit HealthCare (Hefei) Co. Ltd manufactures GastroPanel®-products developed by Biohit Oyj for the Chinese market. Screening studies provided additional information about the diagnosis of gastric cancer risks and the frequency of vitamin B12 deficiency More research evidence was obtained on the association of Helicobacter Pylori with gastric cancer. The long-term follow-up of the subjects who participated in a population-based screening during 1994-1996 was completed and the report was published in Scandinavian Journal of Gastroenterology. The study confirms the previous observations, according to which helicobacter infection alone increases significantly the risk of gastric cancer. This risk increases substantially when an untreated helicobacter infection has progressed to atrophic gastritis. Furthermore, we obtained additional evidence that GastroPanel® test predicts the risk of stomach cancer. In GastroPanel® test, a low Pepsinogen I level predicts stomach cancer risk even ten years in advance. These results were obtained in two separate studies among Asian and Caucasian population, where the subjects were followed-up for several years after GastroPanel® testing. The long-term predictive value of GastroPanel® test as a predictor of this risk has not been previously confirmed in a Caucasian population. According to a study conducted in Estonia and Finland, in the majority of cases in Estonia but less than 25% in Finland, vitamin B12 deficiency remains undiagnosed among elderly people. In that study, the frequency of atrophic gastritis and levels of vitamin B12 were compared among elderly people in both countries. Cancer screening pilots continued in China and Russia We continued our efforts towards advanced medical practices, especially in cancer screenings. The two pilot screening trials for gastric cancer risk started in China in 2015 continued, using Biohit Oyj`s GastroPanel® test. The first of these is a project of the National Clinical Research Center for Digestive Diseases (Changhai Hospital), funded by the Ministry of Science and Technology of China. The Ministry is the coordinator of this multi-center study screening the risks of early gastric cancer. In the study, at least 20,000 persons will be screened in some 50 hospitals. The screenings were expected to be concluded by the end of 2016. The second study is being conducted in Chinese healthcare centers by the China Health Promotion Foundation. The foundation is a public organisation, administered by the Chinese Ministry of Health. Around half a million 40-80-year-old asymptomatic subjects will be screened in this trial. The sample collection has started in the summer of 2015 and continued over the whole year 2016. Due to the fact, that both organisational sides of the study are public organisations, and independent of Biohit Oyj and its joint venture, there is no specific information available about the exact time of completion. In 2015, Russian Federation started a pilot project for colorectal cancer screening, targeted to 48-75-year old asymptomatic persons. In the pilot project, around 20,000 persons will be screened, and the results are expected to be available during 2017. Based on these results, the final selection of the screening test to be used in the national screening program will be made, and one of the options is Biohit Oyj`s ColonViewFIT test. The national screening is organised and sponsored by the Russian Federal Government and it will be conducted by local medical centers. We brought our expertise for food industry In 2016 we started a cooperation with Pyynikin Craft Brewery and brought our biotechnological expertise at the disposal of the brewery production process. As a result of this cooperation, Pyynikin Craft Brewery launched a new beer brand called Pyynikin Vapaa (Pyynikki´s Free), responding to the consumers’ demands. The product is, among other properties, a gluten-free product and it has a very low acetaldehyde level. The cooperation with the brewery is a new business area for Biohit and it paves the way for developing the production of other food stuffs as well. We want to be part of the development aimed to offering the customers alimentary products made of pure raw materials. Biohit's product portfolio consists of diagnostic tests, analysis systems, products binding carcinogenic acetaldehyde into a harmless compound, monoclonal antibodies and service laboratory operations. The entire product and service portfolio is reported under a single segment. NET SALES AND RESULT The operating loss was EUR -3.4 million (EUR -2.9 million 1-12/2015). The result during the reporting period totalled EUR -3.3 million (EUR -2.9 million). On 31 December 2016, the balance sheet total was EUR 13.0 million (EUR 11.7 million). Biohit's balance sheet provides the necessary foundation for building new business and for utilising the significant potential of the company's products. At the end of 2016, our equity ratio was 83.0% (87.9%). Biohit Oyj has a moderate financial position, which allows for the necessary actions towards creating an international distributor network as well as the development and commercialisation of new products. Liquidity is at a good level. At the end of the reporting period, the company's financial assets totaled EUR 7.7 million (EUR 7.2 million) containing EUR 3.2 million worth of Genetic Analysis AS shares. Despite loss making financial periods the company has managed to keep its working capital on a good level and the management believes the working capital will cover the operations for the next 12 months and the company is not dependent on external financing to be able to guarantee the continuity of its operations. Furthermore, as announced on 2 January 2017, the company expects to receive EUR 1.8 million cash during the first half of the year from the ownership arrangement of its joint venture, which is expected to strengthen company’s working capital structure. The transaction requires approval from the authorities. Company’s management assessment is that company’s ability to continue its operations is good and there are no indications towards events or circumstances that alone or together might give a significant reason to doubt the organisation’s ability to continue its operations. R&D operations focus on innovations, as well as product development and improved usability. Biohit also employs external experts and subcontractors in its R&D operations. Development expenditure has not been capitalised. Research and development expenditure during the 1-12/2016 reporting period amounted to EUR 2.0 million (EUR 2.0 million), of which the second half-year accounted for EUR 0.8 million (EUR 1.0 million). The main focus was in the development of the GastroPanel® quick test. The quick test is particularly suitable for the test assortment at doctor’s offices and small laboratories, and it is designed to operate with an automatic reader. During 2016, a dysbiosis test was launched on the market measuring the gut microbiota. A bone density measurement (by a newly designed equipment) was added to the test portfolio of our laboratory services. Ongoing clinical trials continued and new trials were started To confirm the promising results of the smoking intervention study, which was concluded in November, 2015, a new, more comprehensive study with Acetium lozenge was designed. The study setting is similar as in the previous study, in which Acetium lozenge was found to be more effective than placebo in assisting the cessation of smoking. This needs to be confirmed in an adequately powered study. The new study with larger series of patients started in spring 2016 and continues until the first half of 2017. L-cysteine slowly released from Acetium lozenge binds the human carcinogen acetaldehyde in the saliva to form a harmless compound, and in addition, improves oral health. An international study comparing colorectal cancer screening tests still continues in Barretos, Brazil. It has a similar design as the study completed in 2015 but the cohort of screened patients is larger. This study is close to completion and compares the sensitivity and specificity of Biohit Oyj’s ColonViewFIT test and a traditional guaiac-based method as a screening test for colorectal cancer. Based on the interim results, an abstract was submitted for the 2017 congress of DDW (Digestive Disease Week) in the USA. During 2016, the two randomised double-blind trials on patients who suffer from migraine and cluster headaches still continued. Due to the slow recruitment of the patients, the progress of these two studies is delayed. There are no interim results available. Year 2016 was also the starting point for a long-term treatment trial in two clinics in Italy, where the efficacy of Acetium capsule in the treatment of atrophic gastritis or in intervention of disease progression is tested in a randomised placebo-controlled trial. This study setting necessitates a sufficient number of patients who fulfill the selection criteria and an adequate follow-up period after therapy. Due to these reasons, the total duration of the project will be at least three years. The results of the senior citizens study, which was concluded at the end of 2015, were published during 2016 in the Journal of Aging Research and Clinical Practice. In this study, more than 200 residents of assisted housing facilities in Finland (Tampere) and Estonia (Tartu), with an average age of over 80 years, were tested with GastroPanel®- and vitamin B12 measurement. The aim was to compare the prevalence of atrophic gastritis, helicobacter-infection and B12 vitamin deficiency. The study disclosed significant differences in the general health of these elderly people between the two countries. This applies particularly to the prevalence, detection rate and management practices of helicobacter infection and vitamin B12 deficiency. Atrophic gastritis detected by GastroPanel® examination is a common cause of vitamin B12 malabsorption, an early diagnosis of which is emphasised also in this study. Novel results on the use of GastroPanel® test and a meta-analysis In 2016, Biohit’s Department of Clinical Research made a systematic review of the entire literature published on the use of GastroPanel® test in diagnosis of atrophic gastritis worldwide. All studies fulfilling the inclusion criteria were subjected to a formal meta-analysis. Meta-analysis is a statistical technique, which is suitable for calculating the pooled sensitivity and specificity of GastroPanel® test in diagnosis of atrophic gastritis, based on all published studies. Altogether, 27 studies fulfilled the inclusion criteria, comprising nearly 9000 patients tested with GastroPanel® in different countries and continents. Despite substantial variation between individual studies, the pooled results of this meta-analysis are encouraging and clearly substantiate the positive experiences of this test, reported in Biohit´s own studies and in those conducted by its partners. Especially important is the high specificity of GastroPanel® test in detecting atrophic gastritis signifying in practical terms that in cases with a completely normal test result a probability of atrophic gastritis or helicobacter-infection in the stomach is negligible. In addition to this meta-analysis, also two other studies with significant results conducted by Biohit or its partners were reported in Anticancer Research-journal during 2016. One of these is a screening study with GastroPanel® conducted in St. Petersburg with asymptomatic patients and the other one is a case-control study with 10-year longitudinal setting, in which the value of GastroPanel® as a predictor of a long-term risk of gastric cancer was estimated. The suitability of GastroPanel® as a screening tool of asymptomatic population was excellent and the test was sensitive and specific to detect a. o. atrophic gastritis (www.biohithealthcare.com/additional-information ). In the case-control study, the below-threshold levels of GastroPanel® markers (PGI and PGII) were an independent predictor for an increased risk of gastric cancer even during a follow-up of 10 years. In the multi-variate model, the strongest predictor was a low PGI/PGII-ratio, which is an indicator for mucosal atrophy in the GastroPanel® marker profile. During 2016, a new clinical acetaldehyde study in Uppsala University Hospital was concluded. The study compared the effect of Acetium capsule and placebo on the acetaldehyde levels in the gastric juice after alcohol intake in volunteers who had an atrophic gastritis (acid-free stomach) confirmed by GastroPanel® test and biopsy. Like in the previous studies, Acetium reduced the acetaldehyde level in the gastric juice highly significantly (68%) as compared to placebo. Furthermore, the study is the first to confirm that the slow-release L-cysteine remains in the stomach for several hours and binds acetaldehyde into a harmless MTCA-compound. These results were published in the November 2016 issue of Scandinavian Journal of Gastroenterology. Acetium has been in use by Japanese cancer researcher for quite a long time by now. In Akita University the research group of Professor Katsunori Iijima (Head of Gastroenterology Department) has conducted further studies assessing the impact of Acetium on the internal acetaldehyde levels in the oesophageal and gastric mucosa and its possible impact on preventing the recurrence of cancer among the high-risk patients. GastroPanel® examination developed by Biohit Oyj is a significant tool in assessing the cancer risk of these patients. During 2016 the awareness of the harmfulness of acetaldehyde was increased in several occasions. In conjunction with the Annual Medical Assembly professor Mikko Salaspuro held a presentation about acetaldehyde as the most common carcinogen in the world. The presentation provoked a lively public discussion. Professor Salaspuro held a presentation about the same topic also in 8th Annual Charles Lieber Satellite Symposium in New Orleans on 25 June 2016. The presentation was called ‘Acetaldehyde a neglected human carcinogen’ and its summary is published in an international serie ‘Experimental and Molecular Pathology 102: 2017’. In the light of the current knowledge, the packages of alcoholic beverages should contain a statement on its possible content of acetaldehyde, which is classified by the WHO as a human carcinogen. Given that there is no scientific evidence to indicate that acetaldehyde and ethanol in the foodstuffs are less carcinogenic than acetaldehyde in alcoholic beverages, the same requirements should also apply to these products. Several alcoholic beverages and foodstuffs have an acetaldehyde content in excess of 5mg/L, which is the recommended upper limit for the acetaldehyde content in cosmetic products. Additionally, a scientific committee set by the EU has proposed in 2012 that mouthwashes should not contain acetaldehyde at all. Gross investments during the 1-12/2016 reporting period totalled EUR 0.1 million (EUR 0.2 million), of which the second half year accounted for EUR 0.0 million (EUR 0.0 million). Key investments in the period were related to production automation-related equipment. During the review period, the Biohit Group employed 53 (52 in 2015) people on average. 44 (44) of whom were employed by the parent company and 9 (8) by the subsidiaries. At the end of the year 2016, the Group employed 49 (49) personnel, of whom 40 (40) were employed by the parent company and 9 (9) by the subsidiaries. Biohit’s key risks are related to the investments required for business growth and adequacy of economic resources these require in the medium term. Company’s management assessment is that company’s ability to continue its operations is good and there are no indications towards events or circumstances that alone or together might give a significant reason to doubt the organisation’s ability to continue its operations in the next 12-month-period. Other risks are involved in areas such as the success of clinical trials, the selection and development of new market areas and distribution channels, personnel recruitment, registration processes, product pricing, and political decision-making affecting the progress of screening programs. Significant short-term risks are associated with the successful selection of new market areas, the timing of expansion into selected markets and product success in these markets. The recent increase in uncertainty factors associated with international politics may have an unfavorable impact on the company's business. The duration of the product registration process is different in each market area. For this reason, it is not possible to accurately assess the time taken for the authorities to handle registrations in these areas and for product sales to begin. When investing liquid assets, the objective is to gain a return on investment with a minimum risk of equity loss. The investment portfolio consists of deposits, money market investments and corporate loans. A fundamental aspect in portfolio management is sufficient diversification across different asset classes, investment instruments and counterparties. Biohit conducts its investment activities with at least two partners. Biohit’s operation’s customer base is widely diversified, with the exception of GastroPanel® sales in China, which currently represents a major single business for Biohit. Due to this reason, the company is dependent on the continuation of this business relationship. Otherwise, the company is not significantly dependent on individual customers or project deliveries. Most of the company’s business is conducted in euro, and the indirect effects of currency exchange rate fluctuations are considered insignificant. Together with its distributors and license partners Biohit has several product registrations ongoing in a number of different markets, which is affecting net sales development. A number of such registrations are expected to be completed in 2017. In addition, negotiations are in progress with new partners, including on the launch of major screening projects, but a number of political risks are affecting the progress of such projects. China’s operations are in Biohit’s focus in 2017. In addition to the ongoing screening studies Biohit’s joint venture Biohit HealthCare (Hefei) Co. Ltd has succeeded in building a wide distributor network and customer base as well as in commercialising GastroPanel® widely, making use of the previous work on gaining price approval decisions in Chinese provinces. Development of sales has been good and increasing, especially in the second half of the year. The joint venture is well positioned to continue this work also in 2017. Biohit expects to complete the reduction of the joint venture share capital during the first quarter of 2017. As a result of the transaction Biohit Oyj estimates its non-comparable operating result to turn clearly positive for 2017. Reduction of the entire share capital of the joint venture is a part of our future GastroPanel® strategy in China. Establishing a production plant, receiving the needed licenses and commercialisation of the new product required sharing the risk in the means of a joint venture but for a company, which is completely Chinese-owned, is easier to get an access to national health care programs and growth funding than for a company, which is partly foreign-owned. Biohit's cost structure is characterised by high investment in research to obtain further evidence on the efficacy of Biohit's diagnostic tests in various clinical settings and in population-based screenings. Late 2016 Biohit announced GastroPanel® quick test. GastroPanel® quick test differs from the current version by being able to give the result already during a single clinical appointment. GastroPanel® quick test is available in Europe after the performance and clinical tests required by the CE certification process are completed. We also facilitated the access to our health tests. R-kioski convenience stores are selling and marketing gift cards for Biohit Oyj’s health checks for consumers. We aim to grow profitable and are strongly committed to taking necessary actions in order to build a profitable future for the company. Net sales growth is expected in 2017. The company does not assess when the comparable result will turn to positive. The parent company's distributable funds (unrestricted equity) on 31 December 2016 are EUR 5,479,775.77, of which the period net loss is EUR 2,580,940.29. The Board of Directors proposes to the Annual General Meeting that no dividend be paid for the financial year. MAIN EVENTS IN THE SECOND HALF OF THE YEAR (H2) Beijing HuayiHuilang Medical Instrument Co., Ltd. to distribute GA-mapTM Dysbiosis Test in China Biohit Oyj and Beijing HuayiHuilang Medical Instrument Co., Ltd. signed an agreement for the distribution of the GA-mapTM Dysbiosis Test in China. Ilari Patrakka was appointed Biohit Oyj’s Sales and Marketing Director and as a member of the Management Team as of August 26th 2016. He holds a Master of Science (M.Sc.) degree in Business Administration. Biohit Oyj and Glomedics signed an agreement for the distribution of the Biohit Lactose Intolerance Quick Test in South Korea. The first meta-analysis of the scientific studies conducted with GastroPanel® was completed – the test is accurate for diagnosing atrophic gastritis The first meta-analysis of the scientific studies conducted with GastroPanel® test was completed. GastroPanel® is a unique, very informative test for examining patients with upper stomach problems and diagnosing helicobacter gastritis (infection) and atrophic gastritis developed from it and stomach cancer and other risks caused by atrophic gastritis. Atrophic gastritis, which is usually asymptomatic and which 13C urea breath test, fecal antigen test or pure antibody tests can not detect, can be detected only with GastroPanel® or gastroscopy and biopsy examination. Possibly for this reason, a stomach cancer developed from atrophic gastritis is still diagnosed at late stages, when the prognosis is very poor (www.biohithealthcare.com/additional-information ). Biohit Oyj’s Chinese joint venture Biohit HealthCare (Hefei) Co. Ltd production facility passed official test requirements and was granted a license enabling production and sales of products. Biohit HealthCare (Hefei) Co. Ltd will start manufacturing of the globally unique GastroPanel®-product developed by Biohit Oyj for the Chinese market. The first shipments will go to fill orders relating to the ongoing gastric cancer risk screening study. Acetium capsule prevents the exposure to carcinogenic acetaldehyde in patients with atrophy of the stomach mucosa New scientific evidence on Acetium capsule was obtained. A collaborative study with Uppsala University shows that Acetium capsule prevents acetaldehyde exposure in patients suffering from the atrophy of the stomach mucosa (atrophic gastritis) caused by Helicobacter pylori infection. GastroPanel® test’s performance and clinical tests as well as registration can be potentially be completed already during 2017. Thisquick test can be performed during a clinical appointment from a fingertip blood drop. The new test is an advanced version of Biohit´s unique GastroPanel® stomach health test (www.biohithealthcare.com/additional-information). GastroPanel® quick test differs from the current clinical test version by giving the results immediately during a single clinical appointment. The GastroPanel® quick test saves costs and unnecessary clinical appointments as well as speeds up the referral to further examinations and eventual treatment, which significantly improves the patient safety. Biohit Oyj and R-kioski started cooperation. R-kioski will sell and promote the gift cards for Biohit’s health checks for its consumers in R-kioski convenience stores across Finland. The agreement will enter into force immediately and will start with a launch during December 2016. Biohit Oyj’s Board of Directors decided on December 5, 2016 to distribute option rights of the option scheme I 2013 as follows: Graham Johnson 30.000 pcs, Panu Hendolin 30.000 pcs, Niklas Nordström 30.000 pcs, Daniela Söderström 30.000 pcs and Ilari Patrakka 30.000 pcs. For the option rights share subscription period commences on January 1, 2017 and ends on May 31, 2019. The persons have accepted the option rights. Biohit Oyj’s Financial Reporting and Annual General Meeting in 2017 Biohit Oyj will release its financial statements for 2016 on Monday 20th of February 2017.The Annual Report, which contains the Financial Statements for 2016 and the Report of the Board of Directors will be published approximately during week 12. Biohit Oyj’s Annual General Meeting has been planned for Wednesday 26th of April 2017 at 5:00 pm. Half year financial report for period January-June (H1) 2017 will be published on Thursday 17th of August 2017. Possible to send a whole blood sample for GastroPanel® examination Biohit has now subjected the GastroPanel® examination for further refinements, of which a patent application has been recently submitted. Now a whole blood sample can be sent to GastroPanel® examination without additional processing steps in context of drawing the sample, like centrifugation and transfer of the plasma (separated from a whole blood sample) into a tube with patient’s data label. This gives only in Europe the possibility for the thousands of private practices to take GastroPanel® as part of their test assortment with its benefits. Furthermore, by collecting whole blood samples, the risk of stomach and oesophagus cancer, with GastroPanel® screening, which provide plenty of other information as well, could be even easier to realise in a cost efficient way (www.gastropanel.com, www.biohithealthcare.com/additional-information ). MAJOR EVENTS AFTER THE CLOSE OF THE PERIOD Ownership arrangement in Biohit Oyj’s Chinese Joint Venture – 2017 operating result expected to be positive Biohit Oyj and Anhui Wisdom-Win Investment Co. Ltd signed a resolution authorised by shareholders of Biohit HealthCare (Hefei) Co. Ltd. a joint venture operating in Hefei, China, concerning reduction of the joint venture share capital for an amount equal to Biohit Oyj’s shareholding. Biohit Oyj owns 40% of the company, and the agreement is for reduction of the entire share capital. As a result of the transaction Biohit Oyj estimates its operating result to turn calculatory positive for 2017. The transaction requires approval from the authorities. Biohit Oyj does not classify Biohit HealthCare (Hefei) Co. Ltd shares as an asset available for sale as the book value of the asset is not in principal determined by the assets transaction value and the share capital reduction requires approval from the authorities. Biohit's financial reporting and Annual General Meeting in 2017 Biohit Oyj’s Annual General Meeting has been planned for 5.00 pm on Wednesday 26 April 2017 in Helsinki. The Board of Directors will call the General Meeting at a later date. In 2017 the company will publish the half-year report for period January - June 2017 (H1) at 9:30 am on Thursday 17 August 2017. The members of Biohit's Management Team are: CEO Semi Korpela, CFO Niklas Nordström, Director of Business Development Lea Paloheimo, Production & Research and Development Director Panu Hendolin, Sales and Marketing Director Ilari Patrakka, Quality and Regulatory Affairs Director Daniela Söderström and Chief Medical Director Kari Syrjänen. Biohit Oyj’s number of shares is 14,698,533 (14,348,533), of which 2,975,500 (2,975,500) are Series A shares and 11,723,033 (11,373,033) are Series B shares. The Series B shares are quoted on NASDAQ Helsinki in the Small cap/Healthcare group under the code BIOBV. Supposing that the market capitalisation for series A and B shares is equal, the total market capitalisation at the end of the period was EUR 88.9 million (EUR 80.5 million on 31 December 2015). Shares’ trade value during the period amounted to approximately EUR 12 million. At the end of the reporting period on 31 December 2016, the company had 6,402 shareholders (6,594 on 31 December 2015). Private households held 76.17% (78.0%). companies 19.26% (20.1%) and public sector organisations 0.02% (0.0%). Foreign ownership or nominee registrations accounted for 4.41% (1.7%) of shares. Further information on the shares, major shareholders and management shareholdings is available on the company's website at www.biohithealthcare.com/investors . This financial statement release has been prepared in accordance with the requirements of the IAS 34 Interim Financial Reporting standard. Biohit Oyj has applied the same accounting principles in preparing this financial statement release as for its financial statements 2015. The figures in the financial statement release have not been audited. Earnings per share calculated from earnings attributable to equity holders of the parent company STATEMENT OF CHANGES IN EQUITY Consolidated statement of changes in equity on 31 December 2016 Consolidated statement of changes in equity on 31 December 2015 Biohit Oyj’s goods and service sales for our joint venture Biohit Healthcare (Hefei) Co. Ltd increased significantly during the financial period compared to previous year being EUR 3.6 million (EUR 1.4 million in 2015 including the closely associated company Anhui Machinery Development). Biohit Oyj B-shares subscribed with stock options I and II 2013 during the 1-12/2016 reporting period During 2016 no new Biohit Oyj shares were subscribed for with stock options I 2013. As a result of the directed share issue for Genetic Analysis AS in 2016 (350,000 shares), the total number of Biohit Oyj's shares increased to 14,698,533 (14,348,533 shares December 31, 2015) and B-shares to 11,723,033 (11,373,033 shares December 31, 2015). The new shares will correspond to a percentage of 2.38% of Biohit Oyj's total number of shares and 0.49% of the voting rights after registration. The half-year report for January - June 2017 (H1) will be published at 9:30 am local time (EEST) on Thursday 17 August 2017. Biohit Oyj is a globally operating Finnish biotechnology company. Biohit mission is “Innovating for Health” – we produce innovative products and services to promote research and early diagnosis. Biohit is headquartered in Helsinki, Finland, and has subsidiaries in Italy and the UK. Biohit Series B share (BIOBV) is quoted on Nasdaq Helsinki in the Small cap/Healthcare group. www.biohithealthcare.com
News Article | November 21, 2016
Bethesda, MD - November 21, 2016 - The Association for Molecular Pathology (AMP), the premier global, non-profit organization serving molecular diagnostic professionals, today applauds the U.S. Food and Drug Administration (FDA) on its decision to re-evaluate its proposed regulatory guidance for laboratory developed tests or procedures (LDPs). AMP believes this decision is in the best interest of patients, healthcare providers, and the entire field of molecular pathology. AMP has long maintained that the involvement of appropriately trained and qualified laboratory professionals is critical to the development of accurate and reliable LDPs, as well as for correct utilization, precise interpretation, and appropriate application of molecular test results. In October 2015, AMP submitted a detailed proposal to the U.S. Senate Committee on Health, Education, Labor, and Pensions (HELP) that modernizes the current Clinical Laboratory Improvement Amendments (CLIA) regulation program, expands its current network of third-party medical experts, and utilizes scientific expertise from FDA and Centers for Disease Control and Prevention (CDC). The AMP proposal provides assurance of quality, analytical validity, and clinical validity without jeopardizing innovation or patient access to necessary care, and does so in a tiered, risk?based structure that avoids duplication of activities within and between federal agencies. "We are pleased that the FDA has decided not to finalize the guidance and we look forward to our continued discussions and professional collaborations to ultimately develop a streamlined approach that ensures high-quality patient care, enhances transparency, and preserves innovation," said Mary Steele Williams, Executive Director, AMP. "Challenging the FDA's initial draft guidance has been one of our top priorities. I'd like to thank all of our members for their countless hours spent developing and advocating for a CLIA-centric approach to LDP oversight and for their ongoing commitment to putting the patient first and preserving broad access to essential care." For more information on AMP's past position statements and letters on LDPs, please visit: The Association for Molecular Pathology (AMP) was founded in 1995 to provide structure and leadership to the emerging field of molecular diagnostics. AMP's 2,300+ members practice in the various disciplines of molecular diagnostics, including bioinformatics, infectious diseases, inherited conditions and oncology. They include individuals from academic and community medical centers, government, and industry; including pathologist and doctoral scientist laboratory directors; basic and translational scientists; technologists; and trainees. Through the efforts of its Board of Directors, Committees, Working Groups, and members, AMP is the primary resource for expertise, education, and collaboration in one of the fastest growing fields in healthcare. AMP members influence policy and regulation on the national and international levels, ultimately serving to advance innovation in the field and protect patient access to high quality, appropriate testing. For more information, visit http://www. .
News Article | March 16, 2016
All animal studies were performed with the approval of the Institutional Animal Care Committees of Sun Yat-sen University, the University of California San Diego, West China Hospital, and the University of Texas Southwestern Medical Center. The eyeball was enucleated from a one-month-old New Zealand white rabbit and washed with PBS (containing antibiotics) three times. After the cornea and iris were removed, a small cut was made in the posterior capsule of the lens; the capsule with attached epithelium was removed and cut into 1 × 1 mm2 pieces. The pieces of epithelium were cultured in minimum essential media supplemented with 20% FBS, NEAA, and 50 μg ml−1 gentamicin. A 17-week-old human fetal eyeball was purchased from Advanced Bioscience Resources, Inc. (San Francisco, California). Post-mortem human eyes were obtained from San Diego Eye Bank. The human LECs were cultured according to the same methods as above. For in vitro differentiation, LECs were cultured on Matrigel-coated six-well plates or eight-well chambers. Lentoid body was formed after 21 days in minimum essential media supplemented with NEAA, 1% FBS, 100 ng ml−1 FGF2, and 5 μg ml−1 insulin. Images of lentoid tissue were obtained using a Leica M205FA stereo microscope. Membrane-tomato/membrane-green (mTmG)-targeted ROSAmTmG mice were purchased from the Jackson Laboratory (Bar Harbour, ME; stock no. 7576) and maintained as homozygotes. P0-3.9-GFPcre mice expressing an eGFP–Cre recombinase fusion protein under the control of the Pax6 lens ectoderm enhancer and the Pax6 P0 promoter26 were maintained in a FVB/N background. Lineage-tracing experiments were performed by crossing the homozygous ROSAmTmG reporter mouse strain with the P0-3.9-GFPcre deleter strain. Eyes were dissected at P1, P14, and P30 and fixed overnight in 4% formaldehyde. Tissues were then incubated in 10% sucrose and embedded in OCT for cryo-sectioning. Frozen sections were washed in PBS and imaged on a Zeiss Axio Imager fluorescence microscope. Bmi1fl/fl mice were generated as previously described27. Nestin-cre mice28 were obtained from the Jackson Laboratory. For BrdU pulses, mice were injected with 100 mg kg−1 BrdU (Sigma) dissolved in PBS, then maintained on drinking water that contained 1 mg ml−1 BrdU until sacrifice. For gene expression studies, lenses of Pax6P0-3.9-GFPcre mice were dissected under a dissecting microscope. Lens capsular bag was opened from the posterior surface by making three crisscross incisions. The capsular bag was opens and lens material extruded. GFP-positive LECs in the mid-anterior capsular area were separated mechanically from GFP-negative LECs in the remaining capsular areas under a fluorescence microscope. RNA was isolated using RNeasy Mini Kit (Qiagen). To image cataracts, mice were anaesthetized with Avertin, and one drop of 1% Mydriacyl (Alcon) was administered per eye. Eyes were immediately visualized in vivo using a light microscope. For histology, mice were perfused with heparinized saline followed by 4% paraformaldehyde (PFA) in PBS. Dissected eyes were fixed in 4% PFA overnight, embedded in paraffin, and sectioned by the UT Southwestern Molecular Pathology core facility. For BrdU staining, slides were deparaffinized, and subjected to heat-mediated antigen retrieval (in 10 mM sodium citrate, pH 6.0). Slides were stained with primary mouse anti-BrdU (Caltag, MD5000, 1:200) overnight at 4 °C. Slides were subsequently stained with Alexa Fluor 555-conjugated goat anti-mouse IgG1 secondary antibody (Life Technologies, 1:500) and 1 mg ml−1 DAPI (1:500) for 1 h at room temperature. The number of BrdU-labelled cells was divided by the total number of DAPI+ cells in a single layer of LECs. Lentiviral shRNA targeting the human BMI1 gene (NCBI Reference Sequence: NM_005180.8) was purchased from Origene (TL314462), ShRNA targeting sequences were as follow: 5′-AATGCCATATTGGTATATGACATAACAGG-3′ and 5′-GTAAGAATCAGATGGCATTATGCTTGTTG-3′. Two shRNAs were used separately, and a non-effective 29-mer scrambled shRNA was used as a control. Lentiviral shRNA particles were prepared using shRNA lentiviral packaging kit (Origene, TR30022). Viruses were harvested at 48 h and 72 h post-transfection. LECs were cultured on Matrigel-coated 3.5-mm dishes with lentoid formation medium for 30 days. Cells were washed twice with ice-cold PBS, and lysed in RIPA lysis buffer with PMSF. Protein concentration was determined by BCA protein assay kit. Thirty micrograms of total protein lysate was loaded onto 10% SDS–PAGE gel and then transferred to a PVDF membrane (Millipore) at 70 V for 2 h. The membrane was probed with the following primary antibody at 4 °C overnight: anti-αA-crystallin (sc-22389, Santa Cruz), anti-β-crystallin (sc-48335, Santa Cruz), anti-γ-crystallin (sc-22415, Santa Cruz) and anti-β-actin (sc-47778, Santa Cruz), and then incubated with HRP-conjugated anti-rabbit, anti-mouse, or anti-goat secondary antibody for 1 h at room temperature. The immunodetection was visualized using a blot imaging system (Fluor Chem Q, Protein Simple) with ECL buffer (Millipore). New Zealand white rabbits (n = 29, four rabbits died from systemic infections unrelated to surgery. The remaining 25 rabbits were used to assess regeneration), and long-tailed macaques (Macaca fascicularis) monkeys (n = 6) underwent minimally invasive capsulorhexis surgery. Only the left eye of each animal was used for experiments. Slit-lamp biomicroscopy and photography were performed at different time points to monitor lens regeneration. Rabbits were euthanized at day 1, day 7, and one month after surgery, and the treated eyes were enucleated. The lenses were harvested for histologic analysis using haematoxylin and eosin staining. For the macaques, enucleation of the treated eye was performed 4 months post-surgery and the lenses were harvested for the same histologic examinations. The eyes were fixed, paraffin-embedded, and sectioned at 5 μm through the cornea, pupil, and optic nerve with the lens in situ. RNA was isolated from rabbit LECs, mature lens fibre cells and LECs in P0-3.9-GFPcre mice using an RNeasy Mini Kit (Qiagen) and subjected to on-column DNase digestion. cDNA was synthesized using a Superscript III reverse transcriptase kit according to the manufacturer’s instructions (Invitrogen). Quantitative PCR was performed via 40 cycle amplification using gene-specific primers (Supplementary Table 1) and Power SYBR Green PCR Master Mix on a 7500 Real-Time PCR System (Applied Biosystems). Measurements were performed in triplicate and normalized to endogenous GAPDH levels. The relative fold change in expression was calculated using the ΔΔC method (C values <30). Rabbit LECs were fixed in 4% PFA for 20 min, then permeabilized with 0.3% Triton X-100-PBS for 10 min and blocked in PBS solution containing 5% BSA, followed by an overnight incubation in primary antibodies at 4 °C. After three washes in PBS, cells were incubated with secondary antibody for 1 h in room temperature. Cell nuclei were counterstained with DAPI. The following antibodies were used: goat anti-Sox2 polyclonal antibody (Santa Cruz), rabbit anti-PAX6 polyclonal antibody (PRB-278P, Covance), mouse anti-Bmi1 antibody (ab14389, Abcam), and mouse anti-Ki67 monoclonal antibody (550609, BD Sciences). The secondary antibodies, Alexa Fluor 488- or 568-conjugated anti-mouse or anti-rabbit IgG (Invitrogen), were used at a dilution of 1:500. Images were obtained using an Olympus FV1000 confocal microscope. We used BrdU labelling to identify and quantify proliferating LECs from human cadaver eyes. Whole-mount human lens capsules were pulsed with BrdU and then stained with an antibody against BrdU to determine the distribution and density of proliferating LECs. In brief, within 12–24 h after death, lenses from post-mortem donor eyes were obtained from the Eye Bank of Zhongshan Ophthalmic Center in Guangzhou, China. Twelve lenses in total from six donors were used for the experiment. A small puncture injury was made on the anterior surface of a post-mortem human lens using a 30-gauge needle. The lenses were cultured at 37 °C in Dulbecco modified Eagle medium (DMEM) supplemented with 10% FBS in a humidified incubator with 5% CO . The contralateral lens from the same donor was treated under the same conditions but did not receive a puncture injury and was used as a control. To label the proliferating LECs, both groups of lenses were incubated in 100 μg ml−1 BrdU (Sigma-Aldrich) 24 h after the puncture injury. The lens was then removed from the capsular bag, and the lens capsules were fixed in 4% formaldehyde and subjected to BrdU staining using a standard immunohistochemistry protocol according to the manufacturer’s instructions (CST, Boston, Massachusetts). Images were taken using a Carl Zeiss microscope (Jena, Germany). This study was approved by the institutional review board of the Zhongshan Ophthalmic Center (ZOC). Informed written consent was obtained from the parents or guardians of the infants before enrolment, and the tenets of the Declaration of Helsinki were followed throughout the study. The study was conducted in accordance with an international guideline and protocol for visual function measurements in paediatric cataract surgery and a protocol of the Childhood Cataract Program of the Chinese Ministry of Health (CCPMOH) and had an independent data and safety monitoring board of ZOC-CCPMOH. The current standard-of-care treatment for paediatric cataract involves removal of the cataractous lens through a relatively large opening using anterior continuous curvilinear capsulorhexis (ACCC, about 6 mm in diameter, Extended Data Fig. 1), followed by cataract extraction and artificial lens implantation or placement of postoperative aphakic eyeglasses/contact lens in paediatric cataract patients younger than two years. Some patients underwent additional posterior continuous curvilinear capsulorhexis (PCCC) and anterior vitrectomy. We established a new capsulorhexis surgery method to facilitate lens regeneration (Fig. 3a). First, we decreased the size of the capsulorhexis opening to 1.0–1.5 mm in diameter. This results in a minimal wound of about 1.2 mm2 in area, which is only about 4.3% the size of the wound created by the current method. Second, we moved the location of the capsulorhexis to the peripheral area of the lens instead of the central area. A 0.9 mm phacoemulsification probe was used to remove the lens contents and/or cortical opacities. These changes provide significant advantages. First, it considerably reduces the size of the injury, which resulted in a lower incidence of inflammation and much faster healing. Second, it moves the wound scar away from the central visual axis to the periphery, leading to improved visual axis transparency. Third, it preserves a nearly intact transparent lens capsule and layer of LECs, which have regenerative potential and are critically required for the regeneration of a natural lens. The clinical trial is an open label, randomized controlled trial in a study population of paediatric cataract patients (age: 0–2 years). Except the trial participants, all other parties (care providers, outcome assessors) were blinded to treatment allocation. A clinical trial consort flowchart is listed in the Extended Data Fig. 8a. Paediatric patients were enrolled accordingly inclusion and exclusion criteria below (ClinicalTrials.gov identifier: NCT01844258). Inclusion criteria were the following: infants were ≤24 months old, and diagnosed with bilateral uncomplicated congenital cataract with an intact non-fibrotic capsular bag. Exclusion criteria included preoperative intraocular pressure (IOP) >21 mm Hg, premature birth, family history of ocular disease, ocular trauma, or other abnormalities, such as microcornea, persistent hyperplastic primary vitreous, rubella, or Lowe syndrome. In total, twelve paediatric cataract patients (24 eyes) received the new minimally invasive lens surgery (Table 1). Twenty-five paediatric cataract patients (50 eyes in total) were enrolled as the control group to receive the current standard surgical treatment (Extended Data Fig. 8a). Bilateral eye surgeries of the same patient were conducted during the same operation session. We defined the incidence of corneal oedema as a >5% increase in central corneal thickness one week post-surgery, and the incidence of severe anterior chamber inflammation as a Flare value >10 evaluated by Pentacam system (OCULUS, Germany) and Laser flare meter (KOWA FM-600, Japan). Early-onset ocular hypertension was identified as IOP >21 mm Hg by Tonopen (Reichert, Seefeld, Germany) within one month after surgery. Macular oedema was identified by fundus OCT (iVue, Optovue, Germany) as an increase in central macular thickness >10% one week post-surgery. When indicated, VAO, defined by visual decline and the degree to which the fundus was obscured, was treated with YAG laser capsulotomy at follow-up. Compared to infants operated on using our new surgical technique, infants who received the traditional technique had a higher incidence of anterior chamber inflammation one week after surgery, early-onset ocular hypertension, and increased VAO (Table 1). However, in the group treated with our new method, a transparent regenerated biconvex lens was found in 100% of eyes three months after surgery, while no regenerated biconvex lenses formed in the group treated with the standard technique. In addition, 100% of the capsular openings healed within one month after surgery in the experimental group, but no capsular openings healed in the control group. Testing equipment included a set of Teller Acuity Cards (Vistech Consultants, Dayton, Ohio). The set of cards consists of 15 cards with gratings ranging in spatial frequency from 0.32 to 38 cycles per cm, in half-octave steps, and one blank grey card. A 4-mm peephole in each card allows the tester to view the child’s face through the card during testing. Test distance was kept constant by use of an aid to measure the distance from the child’s eyes to the card throughout testing. For 38 cm, the aid was the distance measured from the tester’s elbow to a specific knuckle on the tester’s hand, and for 55 cm, the aid was the length (55 cm) of the Teller Acuity Card. Testers were instructed to hold the cards without wrapping their fingers around the front side of the card, as this may attract the child’s attention. Testers presented the cards directly in front of the child and observed the child either over the top of the card or through the peephole in the card. During each acuity test, a masked visual acuity examiner was aware that the gratings were arranged in order from lower to higher spatial frequencies in half-octave steps, but were masked to the absolute spatial frequency of the grating on each card. The subset of spatial frequencies used for each test was selected according to a pseudorandom order from among three possible subsets of spatial frequencies for the subject’s age group. All three subsets for each age group included spatial frequencies known to be well above the threshold for that age group. To keep the visual acuity examiner masked to the absolute spatial frequency, the visual acuity examiner was not permitted to look at the front of the card to confirm the location of the grating. Instead, the visual acuity examiner asked an assistant to confirm the location of the grating on the card, after the visual acuity examiner had shown a card to the subject enough times to assess whether or not the subject could detect the grating. A clinical examiner was masked to the acuity results and the assigned patient group. Acuity was scored as the spatial frequency of the finest grating and was converted to log values before data analysis. We used a handheld auto-refractometer (PlusoptiX A09, OptiMed, Sydney, Australia) to evaluate the function of the regenerated lenses according to the manufacturer’s methods. Descriptive statistics was provided for the primary and secondary endpoints measured by intervention groups at each time point. Mean and standard deviation was reported for continuous variables and count and percentage is reported for categorical variables. To assess whether the primary outcome, decimal acuity, was significantly improved within each group, we performed the pre-post comparison between decimal acuity measured at baseline and study endpoint using paired t-tests. Normality of the data was checked and non-parametric alternatives, Wilcoxon signed-rank test is considered if the assumption was severely violated. To evaluate whether the mean response profiles in two groups were similar, we used the linear mixed-effect model taking account for the within-subjects correlation. The baseline decimal acuity was not adjusted by the model due to the homogeneity of this measurement as shown in the summary statistics. As the standard-of-care approach requires laser surgery at 3 months while the novel treatment does not, we fit two models using before and after laser surgery data, separately, to demonstrate the superiority of the novel approach. In each model, the outcome is the decimal acuity measured at four time points: baseline, 1 week, 3 months (before or after laser surgery) and 6 months; time (baseline as the reference level), treatment assignment and their interaction are the fixed effects; and patient is the random effect. Significant associations are identified using likelihood ratio test (LRT) by comparing models with and without a fixed effect. A linear mixed-effect model is fit again by dropping out the insignificant fixed effect until the final model is selected. A contrast test is performed when necessary. For the secondary aim, we compared the proportions of each condition of complications between two groups. We assumed the occurrence of complications for eyes from the same patient were independent. The mean difference and its 95% confidence interval was reported. A two-proportion z-test was used with the nonparametric χ2 test as alternative if the normality assumption was violated. All tests were two-sided and a P value less than 0.05 is considered to be statistically significant. Accommodative response was measured by an open-field autorefractor (SRW-5001K; Shin-Nippon, Tokyo, Japan), which allows targets to be viewed at any distance. The paediatric patients were positioned for autorefractor measurement with assistance from their parents. The patients were guided to fixate binocularly at a near target (33 cm, 5 × 5 array of smiley faces of N10 size) and a far target (3 m, 5 × 5 array of smiley faces of N10 size) by a trained and certified investigator or study coordinator. The measurements from non-cycloplegic autorefraction were performed three times at each target distance by the same trained and certified investigator throughout the study, in order to maintain accuracy and consistency throughout the trial. Measurements were taken in the same quiet environment with consistent room illumination to diminish the influence of distracting factors and to maintain subjects’ concentration. The spherical equivalent refractive value (SER) was recorded for each measurement and the mean value was calculated for evaluation of an accommodative response. The value of accommodative response was the difference between SER values for the near and the far target. We also used dynamic retinoscopy to measure the infants’ accommodation29, 30, 31. In brief, we recorded a lens dioptre value using retinoscopy when a patient was guided to fixate on a target 3 m away. Then another lens dioptre value was recorded when the target was moved closer, at a distance of 33 cm from the eyes. The difference between these two measurements was used to evaluate lens accommodative power.
News Article | November 21, 2016
In recognition of wide-ranging contributions to the fields of cancer prevention, patient care, and basic, translational, and clinical research, seven faculty members from The University of Texas MD Anderson Cancer Center have been named Fellows of the American Association for the Advancement of Science (AAAS). Election as a Fellow, a tradition that began in 1874, is an honor bestowed upon AAAS members by their peers. MD Anderson's faculty now includes 40 AAAS Fellows. "It is a great honor to be recognized as an AAAS Fellow, and we are proud to add seven more of our outstanding scientists and clinicians who work tirelessly for our patients and push the frontiers of cancer science," said Ronald A. DePinho, M.D., president of MD Anderson. "As we celebrate 75 years of serving patients around the world, we applaud our faculty members' distinguished work and further commit to Making Cancer History for generations to come." Ralph B. Arlinghaus, Ph.D., professor of Translational Molecular Pathology, for research, teaching and leadership in oncology. He is known for his studies on RNA tumor viruses, the v-Mos oncoprotein and the role of Bcr in Bcr-Abl positive human leukemia. Arlinghaus has authored more than 260 journal publications and book chapters. Powel H. Brown, M.D., Ph.D., chair of Clinical Cancer Prevention, for developing and testing novel therapies for the prevention of breast cancer. Brown is a leader in studies of estrogen receptor-negative breast cancer development and progression. His findings have improved the understanding of breast cancer subtypes and identified critical drug targets for breast cancer treatment. Junjie Chen, Ph.D., chair of Experimental Radiation Oncology, for advancements to the field of DNA damage response and genomic integrity, particularly for clarifying how the breast cancer tumor suppressors BRCA1 and BRCA2 act to promote DNA repair and genomic stability. His work has resulted in a clearer understanding of the complex framework underlying DNA damage and how this might be utilized for cancer therapy. Andy Futreal, Ph.D., chair ad interim of Genomic Medicine, for contributions to the fields of cancer susceptibility genetics and somatic cancer genomics. Futreal pioneered the use of large-scale genomics to characterize and identify novel human cancer genes, leading to the landmark discovery of the role of BRAF mutations in melanoma development and rapid approval of the first effective targeted breakthrough therapy for advanced melanoma. He identified mutations in the BRCA1, BRCA2 and ERRB2 genes that contribute to tumor development, and developed the Catalogue of Somatic Mutations (COSMIC) database of somatically acquired mutations found in human cancer. Eugenie S.Kleinerman, M.D., professor of Pediatrics, for clinical and scientific expertise in osteosarcoma. Kleinerman pioneered the first effective immunotherapy for children with osteosarcoma, improving long-term survival from 70 to 78 percent. She further defined the role of the Fas/FasL, Notch, Osterix and REST proteins in osteosarcoma and Ewing's sarcoma metastasis. Jeffrey N. Myers, M.D., Ph.D., professor of Head and Neck Surgery, for comprehensive genomic characterizations of oral cancers. Myers performed some of the initial analysis of gene expression, copy number, methylation and point mutations in squamous cell carcinomas of the head and neck, oral cavity and skin. He also developed a novel computational approach to classify patients with tumors harboring p52 mutations, which have prognostic and predictive value. David J. Tweardy, M.D., professor of Infectious Disease and division head of Internal Medicine, for distinguished contributions to the understanding of cytokine signaling and the role of STAT proteins in inflammation and cancer development. Using STAT3 inhibitors, Tweardy clarified the contribution of STAT3 to the host damage response. His findings revealed how STAT3 modulation may be harnessed to prevent and treat cancer and chronic inflammatory diseases. Founded in 1874, the AAAS is the world's largest general scientific society. Fellows must be nominated by either the steering groups of the association's 24 sections, or by any three Fellows who are current AAAS members (so long as two of them are not affiliated with the nominee's institution), or by the AAAS chief executive officer. Each steering group then reviews the nominations within its respective section and a final list is forwarded to the AAAS Council, which votes on the aggregate list. This year 391 members have been awarded this honor because of their scientifically or socially distinguished efforts to advance science or its applications. New Fellows will be presented with an official certificate and a gold and blue (representing science and engineering, respectively) rosette pin on February 18 at the 2017 AAAS Annual Meeting in Boston.
News Article | December 14, 2016
Though it has been shown that men who get melanoma are twice as likely to die from the disease as women, the biological explanation for this is poorly understood. Research led by Dr. Alan Spatz, Director of Surgical and Molecular Pathology at the Jewish General Hospital and head of the "X chromosome and cancer" lab at the Lady Davis Institute in Montreal, reveals that the decreased expression of the X chromosome gene PPP2R3B and its protein PR70 are positively linked to tumor progression in this aggressive form of skin cancer. The finding is published in Science Translational Medicine. "We focused our research on the machinery of the X chromosome because we postulated that the inactivation of one of the two X chromosomes in women, as opposed to men who have an X and a Y, and the way this mechanism is regulated, may have deep implications on the cell biology of cancer cells" explains Dr. Spatz, who has a long-standing interest in the role of the X chromosome in cancer. "I proposed in 2004 a theoretical model of X-linked tumor suppressor genes loss and oncogenes activation that since has been validated in many situations. The fact that two X's interact together in females has also implications for cancer. We believe that the genetic specificity of the X chromosome plays a significant function in the gender difference we observe in melanoma. And we see unique regulation of tumor suppressor genes and oncogenes in the X chromosome." Dr. Spatz, Professor of Pathology and Oncology at McGill University, and Dr. Leon van Kempen, COO and Scientific Director Molecular Pathology Center, have extensively studied the gene PPP2R3B, which is located on the X chromosome in females, but on the Y in males. The expression of this gene has been independently correlated with more favourable progression in melanoma and is important because its expression is higher in females. PPP2R3B codes for the PR70 protein, which decreases melanoma growth by negatively interfering with DNA cell replication and, therefore, acting as an X-linked tumor suppressor. PR70 is at the forefront of controlling the cell replication cascade. From a clinical perspective, this research suggests there could be potential anti-cancer therapies in actioning the proteins linked to PR70. The proteomics capabilities that have been developed at the Segal Cancer Centre and Lady Davis Institute at the Jewish General Hospital will eventually be instrumental in profiling and identiying the active proteins that could be most effectively targeted by novel therapies. "I believe this discovery advances our understanding of the specific role of the X chromosome genetics in modulating the expression of genes that are critical in cancer progression," Dr. Spatz said. "Specifically, this is a new avenue for exploring X-linked tumor suppressor genes and oncogenes. I'm confident that we will eventually be able to exploit this discovery to pursue new therapeutic avenues against cancer." "The protein phosphatase 2A regulatory subunit PR70 is a gonosomal melanoma tumor suppressor gene" by Drs. Leon van Kempen, Alan Spatz et al appears in Science Translational Medicine.