Wayne State University

NIH Research Projects · FY 2025 · 2003-05

ABSTRACT The long-term goal of this project is to understand the cellular and molecular mechanisms of adipose tissue establishment, maintenance, and pathophysiological remodeling. Our central hypothesis is that adipose tissue plasticity involves the creation and resolution of dynamic adipogenic niches involving close interactions among specific stromal and immune cell subtypes. While single cell RNA sequencing (scRNA-seq) has identified the stromal and immune phenotypes that are involved in adipose tissue neogenesis, how these cells are recruited to specific locations and interact to create and resolve adipogenic niches has not been addressed. In this continuation we will use cutting edge high-dimension, high-resolution spatial transcriptomics and proteomics to map, order, and test the cellular events underlying adipose tissue neogenesis. Despite recent advances in single cell profiling, the origins of adipocytes and lineage relationships of stromal cells in adipose tissues remains controversial. To address this critical gap in knowledge we will apply newly- developed CRISPR/Cas9-based technologies for unbiased, high-dimension resolution of cell lineages during the life cycle of adipose tissues. This technology will be integrated with complementary techniques of spatial transcriptomics, multiplexed 3-dimensional single molecule fluorescence in situ hybridization and immunofluorescence. This integrated approach will provide comprehensive, unbiased spatial analysis of lineage relationships in postnatal adipose tissue establishment, expansion, aging, and remodeling in vivo. Successful completion of this proposal will provide the first unbiased high-dimension, single-cell resolution map of mouse adipose tissues during establishment, expansion, and aging. We will determine the cellular and molecular mechanisms of adipogenic niche formation and resolution, and resolve long-standing questions of cell lineages in a comprehensive and unbiased fashion.

NIH Research Projects · FY 2025 · 1999-08

PROJECT SUMMARY/ABSTRACT This is a competitive renewal of grant award R01MH59299, enhanced by significant new in vivo multi-modal functional imaging directions (1H fMRS and fMRI). These innovations facilitate a never-before attempted understanding of specific aspects of dysfunctional neurobiology in youth with Obsessive Compulsive Disorder (OCD). OCD is a severe, prevalent, and chronically disabling disorder emerging during childhood/adolescence (80% of cases) with well-delineated clinical phenomenology/nosology. However, the relationship between the clinical phenomenology and modes of brain dysfunction is only generally understood. For example, our prior work using 1H MRS and fMRI has implicated general neurochemistry and function of the dorsal anterior cingulate cortex (dACC). However, dACC engages in contextually dependent excitatory or inhibitory modes of behavior/control that are likely to induce changes in the steady-state excitatory and inhibitory (E/I) synaptic drive of the dACC. However, which of these response modes (and the shift in the E/I balance they induce) is particularly relevant to OCD, and its sensitivity to OCD dimensions (Obsessions or Compulsions) and comorbid anxiety symptoms are completely unknown. Here, we aim to parse apart dysfunction in excitatory and/or inhibitory tone of the dACC in relation to OCD. In doing so, we provide a transformative extension of evidence supported under prior grant iterations demonstrating dysfunctional activation and connectivity of the dACC in OCD youth. Now, we use specifically designed motor paradigms with distinct excitatory or inhibitory response modes which induce complementary demands on dACC function. The tasks are administered during multi- modal functional imaging acquisition that includes ¹H fMRS and fMRI. 1H fMRS, which permits understanding of the functional biochemistry of the dACC, is uncoupled from hemodynamics and is ideally suited to investigate functional imbalances in the E/I synaptic drive of the dACC. In the same participants and using the same tasks, fMRI will be acquired in the service of understanding changes in task-induced whole brain network dynamics and connectomics. This unique project combines the clinical and multi-modal functional neuroimaging expertise at Wayne State University to achieve a transformative explication of the dysfunctional neurobiology of OCD. The combination of ¹H fMRS and fMRI will be acquired in 100 OCD youth and 100 matched healthy controls (12 - 19 years), allowing us to measure: a) brain function via glutamate modulation (¹H fMRS) and the BOLD signal (fMRI); b) brain plasticity related to shifts in the E/I synaptic drive (1H fMRS); and c) network dynamics and connectomics (fMRI). In addition to providing compelling and clinically relevant in vivo characterization of disordered excitatory vs. inhibitory signaling in the pathophysiology of OCD, the proposal provides a scientific blueprint for how multi-modal imaging can explicate in vivo brain function. The accumulated knowledge will promote better diagnostic and treatment approaches through a more detailed characterization of OCD pathophysiology.

NIH Research Projects · FY 2026 · 1997-08

Project Summary/Abstract The Karmanos Cancer Institute (KCI), founded in 1943, is a not-for-profit, cancer-only research and patient care organization in partnership with Wayne State University (WSU) in Michigan. The designation by the NCI as a Comprehensive Cancer Center was achieved in 1978. KCI operates its wholly-owned, free-standing, cancer hospital and ambulatory clinics (KCC) in Detroit and in Farmington Hills, a suburban location. As a member of McLaren Health Care (MHC), a not-for profit health care corporation, KCI is also responsible for cancer research, quality of care, and cancer care operations in all facilities owned and operated by MHC. Our catchment area covers 46 of Michigan's 83 counties with a population of 6.7 million people. This catchment area is home to 95% of our patients, and KCI sees one-third of all new cancer patients in the catchment area. The Southeast region of our catchment area includes the metropolitan Detroit, defined as a tri-county area (Wayne, Oakland, Macomb) and home to 3.86 million residents. The city of Detroit is the largest city in the state (672,000 residents). In 2018, KCI physicians saw approximately 12,000 new cancer patients (including approximately 8,000 analytic cases). The annual number of new cancer cases in these 46 counties is approximately 39,000 and has not changed significantly since 2014. This application profiles the strengths and successes of our four scientific Programs, Tumor Biology and Microenvironment (TBM, 01), Molecular Imaging (MI, 02), Molecular Therapeutics (MT, 03), and Population Studies and Disparities Research (PSDR, 04). The research efforts of these Programs is supported by nine Shared Resources (Cores), including one developing Core. As of December 2019, KCI's 135 scientific members have secured total annual direct project funding of $ 63,162,159 (an increase of 12.3% from 2015) of which $21,864,688 is peer reviewed funding and $11,743,830 is from the NCI.

NIH Research Projects · FY 2025 · 1997-04

Project Summary (Overall) Continuing funds are requested by 18 vision scientists (holding 20 eligible NEI R01 grants) and their personnel to support three resource and an administrative core in the merged Department of Ophthalmology, Visual and Anatomical Sciences (former Ophthalmology and Anatomy/Cell Biology Departments). The Administrative core will support the overall management of the Core. The research cores will enable and enhance vision research at Wayne State University and for two members at nearby (15 minutes) Oakland University. Facilities requested are: lmaging/Histopathology (I/H), Immunology (I) and Tissue Culture/Molecular (TC/M). The I/H core in Scott Hall will provide for confocal laser scanning, light microscopy, immunofluorescence, OCT and other cell imaging analyses (e.g., cell viability and quantification of plasma membrane potential), and train in equipment use. There also is assistance/training in management of digital images, slit lamp photography, and poster and publication production. The I core, at Kresge Eye Institute, will provide immunological assays and consultation to these vision researchers. It will initiate and stimulate innovative research projects that address emerging questions on the immunological basis of many ocular diseases. Among its functions, it will provide for isolation of peripheral blood cells for staining or viable cryopreservation, immunophenotyping, phagocytosis assays, ELISA and Western blot evaluation of cytokines/chemokines and other molecules from tissue lysates and culture supernates, image analysis for Western blots, data management and storage, and access to the Flow cytometers (Acuri 6 and Cytek Northern LIghts), and Cellometer instruments and to the Karmanos Flow facility. The TC/M facility, located in Scott Hall, will assist and train for preparation of specialized media, isolation, purification and characterization of ocular cells for primary culture, subculture and propagation of established cell lines, adherence assays, biofilm evaluation using crystal violet staining, genotyping of mice, cryopreservation of cells, and advice on AAV vector design, DNA cloning, site directed mutagenesis, and DNA transfection. Assistance and training is also available for techniques in molecular biology including PCR array and real time RT-PCR, CRISPR/cas9, storage of bacteria (including multi-drug resistant isolates), vectors and cDNA constructs. Each of the cores is staffed by well-trained research assistants and directors are NEI-R01 supported. The core directors and PI, as a team, prioritize NEl R01 supported studies, promote collaboration, and assist in gathering preliminary data for new NEI R01 submissions.

NIH Research Projects · FY 2024 · 1993-02

ABSTRACT Folates are anionic molecules that cross biological membranes poorly by diffusion. Folate uptake is principally mediated by the reduced folate carrier (RFC; SLC19A1) and the proton-coupled folate transporter (PCFT; SLC46A1). Upon internalization, folates facilitate one-carbon (C1) metabolism, leading to synthesis of glycine, serine and methionine, and purine nucleotides and thymidylate. C1 metabolism encompasses cytosolic and mitochondrial pathways connected by an interchange between serine, glycine and formate. The ubiquitously expressed RFC is the major membrane transporter for folates in cells and tissues. RFC is also an important transporter of clinically used C1 inhibitors (e.g., pemetrexed) for cancer, as well as other indications, and loss of RFC is associated with drug resistance. PCFT mediates folate absorption in the upper gastrointestinal tract. PCFT levels in other tissues are generally modest. Unlike RFC, PCFT transport is optimal at acidic pH, approximating the tumor microenvironment. PCFT is widely expressed in human tumor cell lines and primary specimens. We discovered novel cytotoxic PCFT-targeted C1 inhibitors for cancer and established a comprehensive structure-activity relationship for PCFT that is distinct from RFC. Novel pyrrolopyrimidine compounds (AGF94 & AGF347) showed potent anti-proliferative activities toward PCFT-expressing tumors that were augmented at acid pH. Following internalization, AGF94 inhibited de novo purine (DNP) biosyn- thesis at β-glycinamide ribonucleotide formyltransferase (GARFTase), whereas AGF347 inhibited mito- chondrial C1 metabolism at serine hydroxymethyltransferase 2 (SHMT2), with additional effects on C1 metabolism in the cytosol (DNP biosynthesis at GARFTase and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase, and at SHMT1). Both inhibitors depleted ATP; AGF347 depleted glycine and cytosolic C1 pools, with downstream effects on glutathione and levels of reactive oxygen species, and on mTOR signaling. AGF94 and AGF347 showed promising in vivo efficacies toward early and upstage tumor xeno- grafts. We posit that our PCFT-targeted agents offer an entirely new approach for treating cancer. In this R01 renewal, we explore the unique biology of the facilitative folate transporters and C1 metabolism, with a goal of further optimizing therapeutic applications of our novel agents. We propose in Aim 1 to characterize the cellular pharmacodynamics and molecular regulation of PCFT in relation to PCFT-targeted therapies, including transcriptional mechanisms and the role of protein-protein interactions in regulating PCFT. In Aim 2, we will characterize the cellular pharmacodynamics of mitochondrial C1 inhibitors including their transport and metabolism. An important focus of both Aims 1 and 2 will be on the role of the tumor microenvironment, including the impact of hypoxia and acid pH on anti-tumor drug biology and efficacy of these series. Our proposed studies are distinctive for their novelty and focus on clinical translation.

NIH Research Projects · FY 2025 · 1985-09

Cancer is the 2nd major cause of human mortality in the US. It is our view that an interdisciplinary graduate curriculum with a focus on the biology of cancer that interfaces with clinicians engaged in cancer diagnosis and treatment provides an excellent means of training specialists with a sufficient breadth of perspective for successful careers in cancer research, education, clinical care or policy. This is an application for renewal of a T32 training grant, currently in its 35th year, to support 6 pre-doctoral students in Cancer Biology at Wayne State University (WSU) and the Barbara Ann Karmanos Cancer Institute (KCI). Trainees include PhD and MD/PhD candidates primarily from the Cancer Biology Graduate Program (CBGP) in the Department of Oncology, along with a smaller number of trainees from other affiliated PhD programs at WSU. Training draws from the outstanding clinical and research facilities at WSU/KCI and the expertise of 20 accomplished preceptors in areas including neoplastic development, invasion and metastasis, cancer drug discovery, cancer immunology, and cancer epidemiology. This T32 training grant provides an important cornerstone of our training program in the biology of cancer at WSU/KCI. The CBGP offers specialized didactic training in basic and translational principles of Cancer Biology, along with seminars by nationally/internationally recognized speakers, student-faculty research retreats, and opportunities to attend national/international research conferences. PhD students are introduced to state-of-the-art research methods in molecular and cell biology, chemical biology, pharmacology, immunology, and epidemiology, and to cutting-edge technologies through the KCI research cores, while performing hypothesis-based cancer research. Students “round” with oncologists in the KCI Cancer Hospital and receive didactic training on clinical research by academic physicians who lead our clinical trials. Training in the responsible conduct of research, rigorous experimental design and data science is provided. “Value added” educational opportunities for T32 fellows include a workshop on F30/F31 fellowship preparation, a T32 research mini-symposium, enhanced career networking, specialized technology workshops and research conferences, training in community outreach, and individualized career mentoring. The goals of our T32 training program are to develop scientists with a strong foundational training and perspective in Cancer Biology, with outstanding capacities for critical thinking, effective communication and networking skills needed for professional success in the coming decades. As documented in this application, T32 CA009531 has trained an accomplished group of cancer biologists who have assumed professional positions throughout the United States, where through their educational, research and related efforts, they continue to combat this devastating disease.

Other NSERC · FY 2024

Endocrine disruptors, CRISPR-Cas9, Cellular division, Metabolic health, Adipocyte, Zebrafish, Embryonic development, Developmental toxicity, PFAS, AhR