Daniel Leventhal, MS, PhD

The overall objectives of my dissertation research are to determine the development, antigenic specificity and the functional role of tumor-associated Tregs in the context of prostate cancer. Building off of previous studies in the lab which had identified several Treg-specific T cell receptors (TCRs) which were recurrently enriched in TRAMP prostate tumors, I began investigating the development and antigenic specificities of these cells. These studies, which were carried out in close… Show more

The overall objectives of my dissertation research are to determine the development, antigenic specificity and the functional role of tumor-associated Tregs in the context of prostate cancer. Building off of previous studies in the lab which had identified several Treg-specific T cell receptors (TCRs) which were recurrently enriched in TRAMP prostate tumors, I began investigating the development and antigenic specificities of these cells. These studies, which were carried out in close collaboration with Dr. Sven Malchow, a postdoctoral fellow in the lab, led to several novel findings which were published in Science, the acceptance of abstracts to allow me to present our data at both the AACR and AAI annual meetings, and an AAI Trainee Poster Award. Perhaps one of the most impactful findings was that two of the most highly enriched Treg specificities found in TRAMP prostate tumors were dependent on the expression of peripheral tissue antigens in the thymus by the transcriptional regulator Aire for their development. While Aire had been previously shown to play a critical role in the negative selection of autoreactive T cells against tissue specific antigens, our data challenged current paradigms by showing that it also played a role in Treg development in the thymus. Our data also suggest a potential “division of labor” between Aire-dependent and Aire-independent Tregs in the regulation of autoimmunity to peripheral organs and potentially tumor immune responses, raising several critical areas for investigation of this novel putative Treg subset. Therefore it is the objective of this proposal to elucidate the functional roles of Aire-dependent Tregs in maintaining organ-specific immune tolerance and determine their functional impact on prostate cancer progression. Show less

Core Team member in business plan competition aimed at commercializing technologies held by the National Cancer Institute.
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The Avon Foundation, The Center for Advancing Innovation, and the National Institutes of Health National Cancer Institute have partnered to create the first ever business plan and start-up "Challenge" featuring inventions conceived and developed by scientists at the National Cancer Institute and Avon Foundation grantees to… Show more

Core Team member in business plan competition aimed at commercializing technologies held by the National Cancer Institute.
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The Avon Foundation, The Center for Advancing Innovation, and the National Institutes of Health National Cancer Institute have partnered to create the first ever business plan and start-up "Challenge" featuring inventions conceived and developed by scientists at the National Cancer Institute and Avon Foundation grantees to accelerate commercialization in support of high-growth businesses that spur entrepreneurship Show less

During my second rotational project in the laboratory of Dr. Gajewski I further expanded my expertise in the field of tumor immunology and gained experience utilizing transplantable models of cancer in mice. Previous studies in the lab in which the chemokine expression profiles of human melanoma biopsies were assayed had identified a panel of chemokines exhibiting increased expression correlated with increases in T cell infiltration within the tumor lesions. To elucidate the role of… Show more

During my second rotational project in the laboratory of Dr. Gajewski I further expanded my expertise in the field of tumor immunology and gained experience utilizing transplantable models of cancer in mice. Previous studies in the lab in which the chemokine expression profiles of human melanoma biopsies were assayed had identified a panel of chemokines exhibiting increased expression correlated with increases in T cell infiltration within the tumor lesions. To elucidate the role of tumor-derived chemokines in the regulation of anti-tumor immune responses, the Gajewski lab generated several versions of the transplantable mouse melanoma cell line B16 which expressed the murine homologs of the chemokines identified. It was thus the objective of my rotational project to analyze the effects of each chemokine on tumor growth, immune cell infiltrates and T cell activation/priming against a model antigen co-expressed by B16 tumor cells. This project allowed me to gain new expertise in cancer biology and substantially increased my technical expertise in multi-color flow cytometry.

Techniques Gained: Cell culture, transplantable tumor models, tumor growth analysis, tumor immune cell infiltrate harvest and analysis, IFNγ Elispot assay, advanced flow cytometry and ELISA
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During my first rotational project as a graduate student in the Cancer Biology program at the University of Chicago, I continued to build upon my laboratory expertise and was first introduced to the field of Tumor Immunology. Dr. Schreiber’s laboratory is interested in understanding the dynamics of T cell responses against tumor antigens. One tool the lab wished to utilize in order to track in vivo T cell responses in real time was bioluminescence. However, the sensitivity of commonly used… Show more

During my first rotational project as a graduate student in the Cancer Biology program at the University of Chicago, I continued to build upon my laboratory expertise and was first introduced to the field of Tumor Immunology. Dr. Schreiber’s laboratory is interested in understanding the dynamics of T cell responses against tumor antigens. One tool the lab wished to utilize in order to track in vivo T cell responses in real time was bioluminescence. However, the sensitivity of commonly used Firefly and Renilla luciferases is too low to detect tumor-specific T cell populations in vivo. Therefore, it was the objective of my research project to incorporate a novel luciferase (extracellular Gaussian luciferase), which showed more than 200-fold increased signal, into the current constructs utilized by the lab in order to visualize rare populations of T cells responding to tumor antigens. Due to several technical challenges which arose during my rotational project, I was able to master several critical laboratory techniques and gain critical problem solving skills which would be extremely helpful in future projects. Additionally, throughout my rotation in the Schreiber lab I was able to gain further expertise in the field of immunology, further instilling my interests in the tumor immunology field.

Techniques Gained: Retroviral vector generation, retroviral transduction, primary T cell transduction, flow cytometry, mouse dissection and bioluminescence imaging
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I was first introduced to biomedical research as an undergraduate researcher in Dr. Mark T. Muller’s lab. Choosing to take on an independent research project which had not yet been established in the lab gave me the opportunity to formally design and carry out a study from conceptualization to publication. My project originated from previously published data from a collaborating lab which showed that cancer patients having solid tumors of a variety of tissues types (breast, head and neck… Show more

I was first introduced to biomedical research as an undergraduate researcher in Dr. Mark T. Muller’s lab. Choosing to take on an independent research project which had not yet been established in the lab gave me the opportunity to formally design and carry out a study from conceptualization to publication. My project originated from previously published data from a collaborating lab which showed that cancer patients having solid tumors of a variety of tissues types (breast, head and neck, prostate, etc.) had increased titers of antibodies against the cytoskeletal protein Cytokeratin 8. It was thus my project to understand the dynamics of Cytokeratin 8 utilizing several cancer cell lines and to recapitulate and optimize the previously shown enzyme linked immunosorbent assay (ELISA) diagnostic for increased titers of anti-cytokeratin 8 antibodies in cancer patient’s sera. Throughout the research process I applied for and successfully received over $11,000 in funding through both internal mechanisms in the University and through the American Cancer Society. Additionally, my project resulted in an “Honors in The Major” Thesis and a first author publication in the Undergraduate Research Journal. After graduating, my project was taken on by a collaborating scientist at the MD Anderson Cancer Center of Orlando.

Techniques Gained: Molecular cloning, immunofluorescence, confocal microscopy, basic flow cytometry and ELISA based screen design and calibration Show less