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PITTSBURGH, PA, United States

Fernando Bonetto,Radboud University Nijmegen | Mangala Srinivas,Radboud University Nijmegen | Arend Heerschap,Radboud University Nijmegen | Robbie Mailliard,Celsense, Inc. | And 3 more authors.
International Journal of Cancer | Year: 2011

Monitoring of cell therapeutics in vivo is of major importance to estimate its efficacy. Here, we present a novel intracellular label for 19F magnetic resonance imaging (MRI)-based cell tracking, which allows for noninvasive, longitudinal cell tracking without the use of radioisotopes. A key advantage of 19F MRI is that it allows for absolute quantification of cell numbers directly from the MRI data. The 19F label was tested in primary human monocyte-derived dendritic cells. These cells took up label effectively, resulting in a labeling of 1.7 ± 0.1 3×10 13 19F atoms per cell, with a viability of 80 ± 6%, without the need for electroporation or transfection agents. This results in a minimum detection sensitivity of about 2,000 cells/voxel at 7 T, comparable with gadolinium-labeled cells. Comparison of the detection sensitivity of cells labeled with 19F, iron oxide and gadolinium over typical tissue background showed that unambiguous detection of the 19F-labeled cells was simpler than with the contrast agents. The effect of the 19F agent on cell function was minimal in the context of cell-based vaccines. From these data, we calculate that detection of 30,000 cells in vivo at 3 T with a reasonable signal to noise ratio for 19F images would require less than 30 min with a conventional fast spin echo sequence, given a coil similar to the one used in this study. This is well within acceptable limits for clinical studies, and thus, we conclude that 19F MRI for quantitative cell tracking in a clinical setting has great potential. © 2010 UICC.

Vasudeva K.,Duquesne University | Andersen K.,Duquesne University | Zeyzus-Johns B.,Duquesne University | Hitchens T.K.,Carnegie Mellon University | And 4 more authors.
PLoS ONE | Year: 2014

Chronic neuropathic pain following surgery represents a serious worldwide health problem leading to life-long treatment and the possibility of significant disability. In this study, neuropathic pain was modeled using the chronic constriction injury (CCI). The CCI rats exhibit mechanical hypersensitivity (typical neuropathic pain symptom) to mechanical stimulation of the affected paw 11 days post surgery, at a time when sham surgery animals do not exhibit hypersensitivity. Following a similar time course, TRPV1 gene expression appears to rise with the hypersensitivity to mechanical stimulation. Recent studies have shown that immune cells play a role in the development of neuropathic pain. To further explore the relationship between neuropathic pain and immune cells, we hypothesize that the infiltration of immune cells into the affected sciatic nerve can be monitored in vivo by molecular imaging. To test this hypothesis, an intravenous injection of a novel perfluorocarbon (PFC) nanoemulsion, which is phagocytosed by inflammatory cells (e.g. monocytes and macrophages), was used in a rat CCI model. The nanoemulsion carries two distinct imaging agents, a near-infrared (NIR) lipophilic fluorescence reporter (DiR) and a 19 MRI (magnetic resonance imaging) tracer, PFC. We demonstrate that in live rats, NIR fluorescence is concentrated in the area of the affected sciatic nerve. Furthermore, the 19 MRI signal was observed on the sciatic nerve. Histological examination of the CCI sciatic nerve reveals significant infiltration of CD68 positive macrophages. These results demonstrate that the infiltration of immune cells into the sciatic nerve can be visualized in live animals using these methods © 2014 Vasudeva et al.

Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 273.01K | Year: 2008

DESCRIPTION (provided by applicant): Cellular therapeutics represent great potential for curing and alleviating symptoms associated with debilitating human diseases, such as cancer, cardiovascular disease, and neurological disorders. Currently, no versatil e methodology for tracking cellular products following administration exists, leading to delays in these therapies reaching the clinic. The ability to visualize the trafficking of transferred cells throughout the body non-invasively will give clinicians th e ability to better understand why certain cellular transplants are successful and others are not. In addition, determining the biodistribution of transferred cells is of paramount importance to overcome government regulatory hurdles for these therapies. I n this study, we propose characterizing a novel fluorochemical cell tracking reagent recently created for use with MRI, called Cell Sense. This product is an emulsion of fluorine-containing nanoparticles that is used to label cells in culture and enables t ransferred cells to be visualized with great specificity in vivo using 19F MRI. Cell Sense has been tested extensively in mouse cells and imaged using in vivo rodent models. It has not yet been tested on clinically relevant human cells. In this proposal, w e will examine the Cell Sense labeling characteristics in a variety of primary human immune cell types, including dendritic cells, natural killer cells, and T cells, that are currently being used in immunotherapeutic trials. Also, stem cells, including hum an neural progenitor cells (hNPC) and CD34+ cell types, will be studied. This proposal has two Specific Aims: (1) to develop ex vivo cell labeling protocols and examine the Cell Sense labeling dose in therapeutically relevant human cells; and (2) to determ ine whether the fluorochemical labeling has any detrimental impact on human cell viability or normal function. To achieve these Aims, we will employ an extensive battery of in vitro assays to assess cell loading, cytotoxicity, proliferation, phenotype, and function. Combining these results, we will be able to devise optimal labeling protocols for a diverse range of cell types that are currently being used for human clinical trials. By establishing Cell Sense's ability to label a wide variety of human cells with limited toxicity, we will have achieved a critical milestone in moving closer toward a clinical-grade product that can be used to track current transplanted cells for therapeutic use. We believe that the potential impact of Cell Sense is significant, by removing many of the scientific and regulatory hurdles that developers of cellular therapeutics for the clinic currently face. PUBLIC HEALTH RELEVANCE: Understanding how therapeutically relevant cell types behave following adoptive transfer or transplan tation may help us develop novel therapies. This project examines the use of novel fluorochemical MRI reagents to image and detect labeled, transplanted cells non-invasively. The results of this study will move MRI cell tracking closer to clinical adoption and will provide researchers and physicians the opportunity to selectively follow labeled cells following administration.

The disclosure provides, in part, compositions and methods for producing emulsions. In certain embodiments, emulsions of the disclosure can be used for the detection of inflammation and cell tracking using MRI. The disclosure provides, in part, methods for labeling, detecting and quantifying cell members, in vivo. In certain embodiments, emulsions can be used as an artificial blood substrate.

Ahrens E.T.,University of California at San Diego | Helfer B.M.,Celsense, Inc. | O'Hanlon C.F.,Celsense, Inc. | Schirda C.,University of Pittsburgh
Magnetic Resonance in Medicine | Year: 2014

Purpose: Cellular therapeutics are emerging as a treatment option for a host of serious human diseases. To accelerate clinical translation, noninvasive imaging of cell grafts in clinical trials can potentially be used to assess the initial delivery and behavior of cells.Methods: The use of a perfluorocarbon (PFC) tracer agent for clinical fluorine-19 (19F) MRI cell detection is described. This technology was used to detect immunotherapeutic dendritic cells (DCs) delivered to colorectal adenocarcinoma patients. Autologous DC vaccines were labeled with a PFC MRI agent ex vivo. Patients received DCs intradermally, and 19F spindensity-weighted MRI at 3 Tesla (T) was used to observe cells.Results: Spin-density-weighted 19F images at the injection site displayed DCs as background-free "hot-spot" images. 19F images were acquired in clinically relevant scan times (<10 min). Apparent DC numbers could be quantified in two patients from the 19F hot-spots and were observed to decrease by ∼50% at injection site by 24 h. From 3T phantom studies, the sensitivity limit for DC detection is estimated to be on the order of ∼105 cells/voxel in this study.Conclusion: These results help to establish a clinically applicable means to track a broad range of cell types used in cell therapy. © 2014 The Authors. Magnetic Resonance in Medicine Published by Wiley Periodicals, Inc.

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