Lawrence C.,Karp Family Research Laboratories
Methods in Cell Biology | Year: 2011
The zebrafish has emerged over the past several decades to become a mainstream animal model. While the growth of the use of the fish in basic biomedical research has been characterized by innovation, the methods and tools for fish husbandry, management, and care have been slow to evolve beyond those conceived during the initial establishment of the model system. While these approaches and technologies have certainly served the purposes of the field, they must now be improved to better match the widening scope and scale of research being done in fish. Such advances are made possible by applying new scientific information to the development of more sophisticated approaches for fish husbandry and management, and by considering the lessons learned during the establishment of the rodent model system. This review highlights the recent progress made in various areas of fish husbandry and management and points out new directions for further improvements. © 2011 Elsevier Inc. Source
Zhang Y.,University of Michigan |
Yan W.,University of Michigan |
Collins M.A.,University of Michigan |
Bednar F.,University of Michigan |
And 9 more authors.
Cancer Research | Year: 2013
Pancreatic cancer, one of the deadliest human malignancies, is almost invariably associated with the presence of an oncogenic form of Kras. Mice expressing oncogenic Kras in the pancreas recapitulate the stepwise progression of the human disease. The inflammatory cytokine interleukin (IL)-6 is often expressed by multiple cell types within the tumor microenvironment. Here, we show that IL-6 is required for the maintenance and progression of pancreatic cancer precursor lesions. In fact, the lack of IL-6 completely ablates cancer progression even in presence of oncogenic Kras. Mechanistically, we show that IL-6 synergizes with oncogenic Kras to activate the reactive oxygen species detoxification program downstream of the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling cascade. In addition, IL-6 regulates the inflammatory microenvironment of pancreatic cancer throughout its progression, providing several signals that are essential for carcinogenesis. Thus, IL-6 emerges as a key player at all stages of pancreatic carcinogenesis and a potential therapeutic target. © 2013 AACR. Source
Behren A.,Ludwig Institute for Cancer Research |
Behren A.,University of Heidelberg |
Muhlen S.,Northumbria University |
Muhlen S.,University of Heidelberg |
And 8 more authors.
Oncogene | Year: 2010
The Ras oncogene is known to activate three major MAPK pathways, ERK, JNK, p38 and exert distinct cellular phenotypes, that is, apoptosis and invasion through the Ras-MKK3-p38-signaling cascade. We attempted to identify the molecular targets of this pathway that selectively govern the invasive phenotype. Stable transfection of NIH3T3 fibroblasts with MKK3 act cDNA construct revealed similar p38-dependent in vitro characteristics observed in Ha-Ras EJ-transformed NIH3T3 cells, including enhanced invasiveness and anchorage-independent growth correlating with p38 phosphorylation status. To identify the consensus downstream targets of the Ras-MKK3-p38 cascade involved in invasion, in vitro invasion assays were used to isolate highly invasive cells from both, MKK3 and Ha-Ras EJ transgenic cell lines. Subsequently a genome-wide transcriptome analysis was employed to investigate differentially regulated genes in invasive Ha-Ras EJ-and MKK3 act-transfected NIH3T3 fibroblasts. Using this phenotype-assisted approach combined with system level protein-interaction network analysis, we identified FOXM1, PLK1 and CDK1 to be differentially regulated in invasive Ha-Ras EJ-NIH3T3 and MKK3 act-NIH3T3 cells. Finally, a FOXM1 RNA-knockdown approach revealed its requirement for both invasion and anchorage-independent growth of Ha-Ras EJ-and MKK3 act-NIH3T3 cells. Together, we identified FOXM1 as a key downstream target of Ras and MKK3-induced cellular in vitro invasion and anchorage-independent growth signaling. © 2010 Macmillan Publishers Limited All rights reserved. Source
Banyard J.,Karp Family Research Laboratories |
Banyard J.,Harvard University |
Chung I.,Karp Family Research Laboratories |
Chung I.,Harvard University |
And 9 more authors.
Scientific Reports | Year: 2013
Using an in vivo cycling strategy, we selected metastatic cancer cells from the lymph nodes (LN) of mice bearing orthotopic DU145 human prostate tumors. Repeated rounds of metastatic selection (LN1-LN4) progressively increased the epithelial phenotype, resulting in a new model of tumor cell mesenchymal-epithelial transition (MET). DU145-LN4 showed increased cell-cell adhesions, higher expression of multiple epithelial markers, such as E-cadherin, EpCAM and cytokeratin 18, and reduced expression of mesenchymal markers such as vimentin. The MET in DU145-LN4 cells was accompanied by increased expression of the miR-200 family, and antimiRs to miR-200c and miR-141 induced an EMT. MET also correlated with the loss of miR-424. Ectopic transient and stable miR-424 expression induced EMT, with reduced epithelial marker expression and increased cell scattering. Our model provides evidence for spontaneous MET in vivo. We show that this cellular plasticity can be mediated through the combined action of miR-424 and the miR-200 family. Source
Rao S.,Dana-Farber Cancer Institute |
Rao S.,Karp Family Research Laboratories |
Zhen S.,Dana-Farber Cancer Institute |
Zhen S.,Karp Family Research Laboratories |
And 9 more authors.
Molecular and Cellular Biology | Year: 2010
Murine embryonic stem (ES) cells are defined by continuous self-renewal and pluripotency. A diverse repertoire of protein isoforms arising from alternative splicing is expressed in ES cells without defined biological roles. Sall4, a transcription factor essential for pluripotency, exists as two isoforms (Sall4a and Sall4b). Both isoforms can form homodimers and a heterodimer with each other, and each can interact with Nanog. By genomewide location analysis, we determined that Sall4a and Sall4b have overlapping, but not identical binding sites within the ES cell genome. In addition, Sall4b, but not Sall4a, binds preferentially to highly expressed loci in ES cells. Sall4a and Sall4b binding sites are distinguished by both epigenetic marks at target loci and their clustering with binding sites of other pluripotency factors. When ES cells expressing a single isoform of Sall4 are generated, Sall4b alone could maintain the pluripotent state, although it could not completely suppress all differentiation markers. Sall4a and Sall4b collaborate in maintenance of the pluripotent state but play distinct roles. Our work is novel in establishing such isoform-specific differences in ES cells. Copyright © 2010, American Society for Microbiology. All Rights Reserved. Source