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NIH Research Projects · FY 2024 · 2023-09

Project Summary An important public challenge is how we address the multi-layer complexities associated with individuals who have a cancer diagnosis and cognitive impairment and/or functional deficits. Many organizations including the American Society of Clinical Oncology and the National Academy of Medicine (formerly Institute of Medicine) have recognized the need for more research and education about the diagnosis and management of geriatric syndromes in patients with cancer as many healthcare providers (HCPs) are unprepared. The Geriatric Oncology Cognition and Communication (Geri-Onc CC) training program has been designed to train oncologists, primary care providers, and other HCPs (nurses, nurse practitioners, physician assistants, social workers, physical therapists, occupational therapists, etc.) to screen for and conduct an initial assessment to identify cognitive impairment and/or functional decline in the older cancer patient, to learn about factors to consider when treating older cancer patients, and to improve communication with the patient and his/her caregiver. The first 5 years of Geri-Onc CC have been highly successful in terms of the ability to recruit HCP’s from a variety of clinical settings and participants’ ratings of the program, their increased knowledge and self-efficacy, and demonstrated improvements in their communication skills. In this renewal, we plan to build on this success and continue to train the workforce in how to work with older adult cancer patients. Two hundred eighty-eight HCPs working with older oncology patients will participate in the interactive, multi-modal two-day program with ongoing professional development over a 12-month period. Training will occur in 12 cohorts over the 5-year project. We will examine knowledge, attitudes, and use of skills before and these variables as well as uptake and use of communication skills and transfer of learning and skills to the HCPs’ clinical setting up to 12 months after the training. The overarching goal of this cancer educational program is to prepare the workforce to care for this growing, vulnerable, aging population.

NIH Research Projects · FY 2025 · 2023-09

The goal of the Medical Students Summer in Oncology at Anderson Research (Med Students SOAR) program is to provide 25 talented first-year medical students per year with an individualized, hands-on research experience in oncology under the mentorship of our expert faculty, while exposing participants to multidisciplinary cancer care through clinical observations and interprofessional simulation trainings. An increase in knowledge of all areas of oncology will be included in the curricula, as well as career development and networking opportunities. The program objective is to promote careers in cancer research, which is in line with the NCI’s mission to enhance the training of a workforce to meet the nation’s biomedical, behavioral, and clinical research needs. This objective will be accomplished through a formalized 8- to 10-week program at The University of Texas MD Anderson Cancer Center in Houston. MD Anderson, an NCI-designated cancer center that is widely recognized for its outstanding clinical care and number one ranking in awarded NCI grants, is ideally positioned to provide this research experience. Our highly regarded clinical care combined with an emphasis on research makes it an environment unparalleled for mentored research and educational programs. The Program Directors (Marites Melancon, PhD; Vickie Shannon, MD; and Jillian Gunther, MD, PhD) are faculty members with successful research programs and experience in leading education programs. Participating faculty members are not only committed educators but also internationally recognized experts in various fields, spanning basic, translational, and clinical research. Students will be matched with faculty mentors, and they will discuss and decide on the research project prior to the summer. During the summer session, students will complete their projects with structured mentoring and evaluation. Instruction in the Responsible Conduct of Research will be included. Students will also learn about other areas of cancer biology and career development through program lecture series, clinical observation, interprofessional clinical simulation, near-peer mentoring, communication skills workshops, and seminars. The program will culminate in a Summer Experience Final Exposition, in which students present their research project to faculty, students, and staff. Our recruitment plan seeks to attract highly qualified applicants based on academic performance and standing; demonstrated research interest and experience; clarity, focus, and originality of the personal statement; relevance of stated career goals to oncology research; and the strength and relevance of letters of recommendation. Successful implementation of this program will directly address the need to cultivate interest in oncology, and specifically cancer research, early in medical careers in order to recruit and retain future cancer researchers.

NIH Research Projects · FY 2025 · 2023-09

Project Summary There remains a need to develop biomarker tests for personalized risk assessment of breast and ovarian cancers. Such tests would not replace screening programs but would instead be a basic tool that can be integrated with other risk models based on a subject’s characteristics to personalize the risk of harboring cancer and inform on the need for screening and surveillance for earlier detection. The primary translational objective of this proposal is to develop a multi-analyte blood-based biomarker panel based on circulating proteins and autoantibodies against tumor antigens that inform about a subject’s probability of harboring a breast or ovarian cancer. Studies by the applicant team have led to the identification of a panel of cancer-relevant circulating proteins as well as autoantibodies against tumor proteins, including TP53 and novel citrullinated antigens, for detection of breast and ovarian cancers. The PLCO cohort is an excellent resource to further advance testing of candidate biomarkers and to also establish combination rules together with subject characteristics for individualized risk assessment of breast and ovarian cancers to optimized screening and surveillance for earlier detection of these diseases. In Specific Aim 1, leveraging pre-diagnostic plasmas from 969 breast cancer cases and 106 ovarian cancer cases as well as four times the number of non-case plasmas from female PLCO participants that did not develop cancer during study follow-up, we will assess the time-dependent (e.g. 0-1 year, 1-2 years, etc) predictive performance (AUC, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV)) of candidate biomarkers for detection of breast and ovarian cancers. Priority biomarker candidates will be advanced to establish models together with pertinent patient characteristics (e.g. Gail Model for breast cancer) for 1-year risk prediction of BrCa and OvCa. For modeling, we will adhere to the Predictability, Computability and Stability framework. The entire PLCO specimen set will be divided into a Development Set and a Set-Aside Test Set. Modeling and tuning of hyperparameters as well as initial validation will be performed in the Development Set. The model with the best predictive performance (AUC) in the Development Set will be selected for subsequent testing in the Set-Aside Test Set. In Specific Aim 2, we will leverage serial samples procured from cases preceding a diagnosis of a BrCa or OvCa and serial samples for non-cases, and we will test whether longitudinal trajectories of biomarker panel scores improve risk prediction. The proposed study represents a validation of cancer-associated protein and autoantibody biomarkers and has high probability to develop a blood test that can be implemented in the clinical setting for individualized risk assessment of breast and ovarian cancers.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY or ABSTRACT I am a tenure-track Assistant Professor at UT MD Anderson Cancer Center. I have expertise in health disparities, community engagement, and community-based participatory research in the areas of cancer health disparities, cancer survivorship, and clinic-based interventions. However, I have not developed, implemented, or analyzed a multi-level community-based intervention for breast cancer survivors. Under-resourced people face disparities in all stages of the cancer continuum, and their survivorship needs remain unmet. Current research on endocrine disrupting chemicals (EDCs) in hair products indicate an increased risk and recurrence of breast cancer for these communities. There is also a dearth of psychosocial interventions that address the unique needs of breast cancer survivors. Behavioral interventions for this population lack content on cultural backgrounds, hair care, and harmful environmental exposures. Interventions are urgently needed for breast cancer survivors to address their unique survivorship journeys and reduce harmful environmental exposures during survivorship. My goals are to develop, implement and disseminate community-based interventions that improve quality of life (QOL) and address health behaviors related to backgrounds that contribute to disparities affecting cancer risk and survivorship. As a first step towards these broad goals, with this K01 proposal, I seek to develop and evaluate an intervention to improve QOL and reduce adverse chemical exposures of EDCs found in personal care products of breast cancer survivors, post-treatment. The proposed career development and training plan support my trajectory toward becoming an independent, community-based implementation scientist with a focus on cancer health disparities through training and experience in 1) Designing community-based multi-level interventions; 2) Assessing environmental exposures in a community-based multi-level intervention; and 3) Using implementation science to evaluate community-based interventions for under-resourced communities. This project will take place at UT MD Anderson Cancer Center with mentors who are experts in training and mentoring K01 trainees and assessing the psychological, biological and social influences on health outcomes (Primary: Laura Glynn, PhD, Chapman University); randomized intervention designs (Co-Mentors: Virginia Sun, PhD, RN, COH; Lorna McNeill, PhD, MPH, MD Anderson); clinical implications of intervention design for breast cancer survivors (Co-Mentor: Abenaa Brewster, MD, MHS, MD Anderson); environmental exposures (Advisor: Robin Dodson, ScD, Silent Spring Institute); and dissemination and implementation science (Co-Mentor: Robert Newton, PhD, Pennington Biomedical).

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY All stem cells have the capacity for self-renewal, an ability to create daughter stem cells without differentiating into other cell types. Stem cells receive the signals from their niches that instruct them to self-renew and prevent them from differentiating through a cascade of inter-organelle communication processes. Substantial evidence has recently revealed that glioblastoma, the most common and lethal type of brain tumor, has “roots” in a population of glioma stem cells (GSCs) that possess an inexhaustible ability to self-renew. Unlike neural stem cells (NSCs), GSCs are also able to sustain their stemness in the suboptimal environments they encounter outside their niches during invasion. How GSCs, but not NSCs, are able to maintain their stemness outside the niches remains unclear. To identify potential glioma suppressors that affect interaction of GSCs and niches, we discovered that RNA-binding protein Quaking (QKI) is a key regulator of endocytosis that controls receptor trafficking, degradation, and signaling desensitization. Mechanistically, QKI regulates pre-mRNA stability of genes that regulate lipid components of endolysosomes, particularly the unsaturated fatty acids (UFAs). As a consequence of defective endolysosomal function, we showed that depletion of QKI and inhibition of UFA biosynthesis led to the enrichment of cytoplasmic membrane-bound receptors that are required for maintaining stemness. In addition, since polyunsaturated fatty acids (PUFAs) are the substrates of ferroptosis, downregulation of PUFA due to Qki loss renders GSCs resistant to ferroptosis, a major tumor suppression mechanism. Supporting the importance of intracellular vesicle trafficking system regulated by QKI and UFA biosynthesis in glioblastoma, we found that lower levels of QKI, endolysosome and Stearoyl-CoA desaturase (SCD, the key enzyme for UFA biosynthesis) all correlate significantly with poorer prognosis in glioblastoma patients. Our long-term goal is to develop therapies that target the defective endolysosome function in glioblastoma. Given that QKI is a major regulator of SCD genes and inhibition of both QKI and UFA biosynthesis can impair the endolysosome activity and promote gliomagenesis, we hypothesize that QKI deletions/mutations disrupt endolysosomal function in NSCs and GSCs through downregulation of UFA biosynthesis and restoration of PUFA levels can sensitize tumor cells to ferroptosis. To test this hypothesis, we will (a) determine the role of Scd1/2-mediated UFA biosynthesis in QKI-regulated endolysosome functions in both NSCs and GSCs, (b) clarify the mechanism by which QKI regulates Scd1/2 pre-mRNA stability in both NSCs and GSCs, and (c) evaluate the effects of restoration of PUFA levels in sensitizing tumor cells to ferroptosis. Together, these studies will elucidate the molecular mechanisms of how the glioma suppressor QKI regulates intracellular vesicle trafficking in NSCs and GSCs through lipid metabolism, and more importantly, they will contribute to the development of therapeutic strategies that specifically target QKI/SCD-depleted glioblastoma.

NIH Research Projects · FY 2026 · 2023-08

PROJECT SUMMARY/ABSTRACT Genomic imprinting is an epigenetic gene-marking phenomenon that causes a subset of mammalian genes (i.e. imprinted genes) to be expressed from only one of the two parental copies. The majority of imprinted genes are arranged in chromosomal clusters. Each cluster has an imprinting control region (ICR) with one or more differentially methylated regions (DMRs) - regions marked by DNA methylation on only one allele - which act as the epigenetic signal that controls monoallelic expression. Imprinted DMRs are classified as germline (primary) DMRs (gDMRs) and somatic (secondary) DMRs (sDMRs). gDMRs are established during gametogenesis, resulting from differential de novo methylation of male and female germ cells. sDMRs represents allele-specific methylation acquired during embryonic development. Genetic studies have demonstrated that the de novo DNA methyltransferase DNMT3A is responsible for the establishment of methylation imprints at gDMRs during germ cell development and that the maintenance enzyme DNMT1 is responsible for maintaining methylation imprints during embryonic development and in adult tissues. Despite these advances, fundamental questions remain to be answered, e.g. How sexually dimorphic patterns of DNA methylation in gametes are regulated? And how gDNAs control sDMRs? Preliminary studies in the applicant’s laboratory showed that conditional ablation of the arginine methyltransferase PRMT7 in male germ cells results in incomplete methylation of the gDMR at the H19-Igf2 imprinted locus in sperm. As a result, the progeny show biallelic repression of Igf2 and allelic switch in H19 and Gtl2 expression (Gtl2 is also a paternally imprinted gene). The applicant’s laboratory also showed that PRMT7 catalyzes monomethylation of DNMT3A at a conserved arginine residue in the N-terminal region that is important for functional specificity of DNMT3A. The applicant hypothesizes that PRMT7, through methylating DNMT3A, regulates DNMT3A-mediated de novo methylation, including the establishment of paternal imprints, during male germ cell development and that, in the absence of PRMT7, impaired DNA methylation on the paternal alleles causes secondary trans-effect changes on the maternal alleles after fertilization, resulting in allelic switch in the expression of some paternally imprinted genes. To test the hypothesis, the applicant proposes two specific aims: 1) Determine the role of PRMT7 in de novo DNA methylation during male germ cell development; and 2) Elucidate the mechanism underlying allelic switch of paternally imprinted genes in the offspring of PRMT7-deficient male mice. In the applicant’s opinion, the proposed research is innovative for its conceptual novelty. The project is significant, because results from the proposed studies are expected to provide novel insights into the regulation of de novo DNA methylation in the male germline and the crosstalk between the paternal and maternal alleles to control monoallelic expression of imprinted genes.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Cellular senescence is a tumor-suppressive cell growth arrest triggered by inducers such as stand-of-care epithelial ovarian cancer (EOC) chemotherapeutic platinum, known as therapy-induced senescence. However, senescent cells are viable and may promote therapy relapse and immune escape through the secretion of factors such as cytokines, chemokines, and growth factors, termed the senescence- associated secretory phenotype (SASP). Thus, it would be ideal to selectively eliminate the detrimental SASP while maintaining the senescence-associated growth arrest. Developing novel therapeutic strategies to overcome therapy resistance remains a major obstacle to overcome in combating EOC. Thus, the overall goal of this proposal is to investigate the mechanism underlying the SASP and leverage these newly gained mechanistic insights to develop senescence based combinatory EOC therapeutics. cGAS promotes the SASP through recognizing cytoplasmic chromatin fragments (CCF) during senescence. Our preliminary studies show that the protein Thioredoxin Reductase 1 (TXNRD1) is localized to CCF and TXNRD1 inhibition impairs the localization of cGAS into CCF, the cGAS-STING pathway, and the SASP during platinum-induced senescence in EOC. Notably, TXNRD1 inhibition does not affect senescence- associated growth arrest. The objectives of this application are to investigate the signaling basis by which TXNRD1 controls the SASP and to investigate a combination senescence based EOC therapeutic strategy. Our central hypothesis is that TXNRD1 promotes therapy relapse and resistance through the SASP by activating the cGAS-STING signaling pathway during therapy-induced senescence in EOC. Accordingly, two specific aims are proposed: Aim 1 is to elucidate the molecular mechanism by which TXNRD1 regulates the SASP during senescence, and Aim 2 will determine the role of TXNRD1 in EOC therapy response. The proposed studies are highly novel because this is the first study to explore a molecular switch that controls the SASP by regulating the cGAS-STING signaling pathway via CCF. Thus, our studies are paradigm-shifting in their potential to elucidate the molecular basis of SASP regulation during senescence. The research proposed is of high impact because it will lay the critical foundation for ultimately developing urgently novel EOC therapeutic strategies through limiting SASP-associated therapy relapse and resistance. Therefore, the current study will not only provide critical mechanistic insights into SASP regulation during senescence but will also have far-reaching implications for the development of senescence-based therapeutic strategies.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Chimeric antigen receptor (CAR) T cell therapy has revolutionized the treatment of hematologic cancers. However, for solid tumors, CAR T cells face challenges including intratumor heterogeneity, dynamic expression of target receptors, and often the inability for T cells to traffic to tumors to mediate the desired antitumor effect. In contrast to the lack of T cell infiltrates, many solid tumors are abundant in immune suppressive myeloid cells including macrophages. Therefore, converting these immune suppressive cells into tumoricidal phenotype represents a promising strategy for cell-based therapy. There are now strong interest in generating CAR macrophages in which autologous macrophages are transduced with CAR delivered by viral vectors ex vivo to enhance their phagocytosis, antigen presentation and cytokine producing capabilities following re-infusion. However, ex vivo preparation of CAR macrophages is complex, time consuming, and due to the non-dividing nature of macrophages, is often inefficient. With the recent advances in mRNA-based therapeutics, it is now possible to reprogram specific immune cell populations in vivo, thus eliminating the complex ex vivo production of autologous CAR cells. Our present proposal aims to propose an innovative strategy of generating CAR macrophages in vivo using mRNA-loaded exosomes to treat HER2 receptor positive breast cancer. This will be the first study to evaluate the feasibility of producing CAR macrophages in vivo using mRNA delivery platforms and assessing the antitumor efficacy of CAR macrophages for cancer immunotherapy. We hypothesize that our strategy represents a revolutionary way to produce CAR macrophages in vivo using CAR mRNA-loaded exosome and offers a promising new approach for cell therapy against solid tumors. Our previous study showed that we can efficiently produce mRNA-loaded exosomes to restore protein expression in solid tumors. Furthermore, our preliminary experiments showed that the exosomes loaded with HER2 CAR mRNA can produce CAR macrophages in vivo with enhanced effector functions. Our current study will test our overall hypothesis by using the following specific aims. In Aim 1, we will evaluate the dynamics and toxicity of CAR macrophage production in vivo using CAR mRNA exosomes. In Aim 2, we will evaluate transcriptomic and functional profiles of in vivo generated CAR macrophages, Finally, in Aim 3, we will assess the antitumor effect of in vivo generated CAR macrophages against both murine and human HER2 expressing breast cancer. If successful, our proposed research can overcome a major technical hurdle that is currently facing cell therapy. The mRNA exosome platform could potentially be expanded to other CAR constructs and greatly expand the potential utility of cell therapy for breast and other solid cancers.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Hospitals are increasingly consolidating into health systems with shared ownership and management. Care in a health system has potential benefits for surgical cancer patients including improved access, care coordination, and strategies to disseminate and implement a rapidly evolving evidence-base into practice across system hub and spoke sites. Prior research has demonstrated that these potential benefits remain elusive. The impact of consolidation on quality varies widely, and there are disparate outcomes for surgical cancer patients treated at different facilities in the same systems. We have shown that location of surgical cancer care determines whether effective treatments are adopted (implementation) or ineffective treatments discontinued (de- implementation). We hypothesize that health system characteristics and strategies are associated with variability in implementation of oncologic evidence among hub and spoke hospitals and that through exploration of observed differences we will identify levers for targeted, multi-level interventions. This work addresses the NIH Blueprint objective to enhance research investments by ensuring adoption into practice and targets rural individuals, a population with disparate healthcare access and outcomes, who are often treated at spoke hospitals. We will examine the influence of treatment for common cancers in health system hubs and spokes on patient access and receipt of evidence-based care by linking SEER-Medicare data with health system data. Then, we will identify health system characteristics associated with evidence implementation, both quantitatively using multilevel modeling and qualitatively through structured interviews with health system stakeholders. Finally, we will use the resources within our Health System at the University of Alabama at Birmingham (UAB) to develop a system-level intervention for dissemination and implementation of oncologic evidence across hub and spoke sites. My long-term goal is to become an independent investigator who improves the quality of cancer care delivery by designing, implementing and studying health system-level interventions to increase clinical adoption of oncologic evidence. Through this training award, I will complement my health services and quality improvement science expertise with advanced training in the organization of healthcare delivery, multilevel analysis of secondary data, and implementation science to develop a system-level intervention to improve evidence-based surgical cancer care.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT The overall vision of the AVANÇO Research Consortium is to establish sustainable infrastructure to develop research programs focused on HIV-associated malignancies in Brazil and Mozambique. The research theme chosen for this U54 HIV-Associated Malignancy Research Center (HAMRC) is to develop novel, low-cost and technologically feasible approaches for cervical cancer screening, diagnosis, and treatment in women living with HIV (WLWH) in low- and middle-income countries (LMICs). This HAMRC will be a collaboration between institutions in the US, Mozambique and Brazil and includes investigators with expertise in HIV/AIDS, Cervical Cancer, Bioengineering, Pathology, Epidemiology and Behavioral Science. The investigators have several ongoing scientific and clinical collaborations which include projects across all three countries. Our proposal addresses the NIH priority: To address HIV-associated comorbidities, coinfections and complications, which includes Malignancies as a category. HIV infection is a known risk factor for cervical cancer, with WLWH having an approximate 6-fold increased risk of developing cervical cancer. The overall goals of the AVANÇO Research Consortium include: 1) Establish sustainable research infrastructure to perform multi-institutional research studies related to HIV-associated malignancies across institutions in the US, Brazil and Mozambique. We will build a comprehensive research program focused on identifying novel, low-cost and technologically feasible approaches for the prevention, diagnosis, and treatment of cervical cancer in WLWH in LMICs. 2) Conduct two Research Projects that advance the development of promising low-cost technologies focused on the diagnoses of high-grade cervical dysplasia and invasive cervical cancer among WLWH. 3) Facilitate and enhance the professional development of junior investigators to conduct HIV- associated malignancy research in Mozambique and Brazil. Through our Developmental Core and Shared Resource Core, we will build research capacity and collaboration through formal research training courses, pilot project funding, and implementation of a Project ECHO® program for research capacity building focused on mentoring and supporting early-career investigators.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Overexpression of epidermal growth factor receptor (EGFR), which frequently occurs in head and neck squamous cell carcinoma (HNSCC), correlates with poor patient survival. However, therapies targeting EGFR in multimodality therapy for HNSCC have not significantly improved outcomes for advanced stage disease. Therefore, alternative approaches targeting EGFR and associated critical pathways are needed to combat HNSCC, the sixth most diagnosed cancer worldwide. Our previous research identified microRNA-27a* (miR- 27a*; miR-27a-5p) as a regulator of EGFR, protein kinase B (AKT1), and mammalian target of rapamycin (mTOR). All these proteins are commonly upregulated in cancer cells, likely as a consequence of tumors repressing miR-27a* expression, and provide a cell survival advantage. Furthermore, re-introduction of miR- 27a* into tumor cells in vitro and in vivo causes apoptosis, raising the exciting prospect that by simultaneously targeting multiple oncogenic pathways, miR-27a* may be an effective therapy for HNSCC. Accordingly, our long-term clinical translational objective is to develop miR-27a* as an effective multimodality therapeutic option for HNSCC, which is directly relevant to the mission of the NIDCR “…to improve oral, dental, and craniofacial health through research...”. This requires us to understand the functional role of miR-27a* targets, to define novel therapeutic combinations that enhance the ability of miR-27a* to inhibit HNSCC progression, and to develop an approach to translate miR-27a* into the clinical arena. Currently, a knowledge gap exists regarding validated targets of miR-27a* and the pathways they influence in HNSCC progression, as well as combinatorial treatments that could augment miR-27a* anti-tumor effects. Moreover, methods for tumor-specific delivery of miRs are lacking. Accordingly, we will comprehensively test the potential of miR-27a* in conjunction with established and novel combinatorial agents for HNSCC treatment using in vitro and orthotopic in vivo tumor models. To overcome challenges in the delivery of miRs in vivo, we will use a novel ultrasound-targeted microbubble delivery platform to administer miR-27a* specifically to tumor. The overall objective of this proposal is to determine the role of miR-27a* in modulating biological processes through regulation of its target genes, while leveraging these and previous findings towards the therapeutic use of miR-27a* within the context of current standard of care and future combinatorial treatment regimens. Our central hypothesis is that re- introduction of miR-27a* in HNSCC negatively modulates critical oncogenic drivers to promote tumor apoptosis. We propose three Specific Aims: (1) To confirm direct molecular targets of miR-27a* that mediate HNSCC pathogenesis; (2) To define miR-27a*-combinatorial treatment regimens for HNSCC; and (3) To characterize the role of miR-27a* delivery in enhancing current and future multimodality treatment regimens for HNSCC. The findings will significantly impact our understanding of miR-27a* targets, the processes they regulate, their role in tumor progression, and ultimately inform clinical implementation of miR-27a* therapy.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Immune checkpoint blockade (ICB) targeting CTLA-4 or PD-1 rapidly assumed its role as a standard treatment for solid tumors and can lead to dramatic, long-lasting responses; nonetheless, fewer than 30% of patients respond to monotherapy with either agent. Combination therapy results in better long-term survival outcomes, but also causes more frequent and severe immune-related adverse events (irAEs). Several novel immunotherapies are currently being explored to evaluate their anti-tumor capacity. Crucially however, most of these targets suffer from on-target, off-cell effects, as other immune cell types can express high levels of these molecules. Hence, as low overall response rates, off-cell effects and widespread immune related toxicity severely limit both treatment efficacy and monotherapy and combination therapy options, there is urgent need to develop novel immunotherapy targets that exhibit a more restricted expression profile. We have recently demonstrated that PD-1+ follicular regulatory T (TFR) cells were prevalent in tumor tissues of several cancer types in humans and mice, and that they were critical cellular determinants of anti-PD-1 treatment efficacy. TFR cells were primarily located within tertiary lymphoid structures (TLS) and exhibited superior suppressive capacity and in vivo persistence when compared to regulatory T (TREG) cells, suggesting a key role for TFR cells in impairing patient survival and impeding immunotherapy treatment efficacy. While we have shown that intratumoral TFR cells derive from TREG precursor cells, the mechanisms and transcription factors (TFs) that are driving this differentiation step are largely unknown. In Aim1, we propose to employ single-cell RNA-seq, single-cell ATAC-seq and micro- scaled ChIP assays to fully characterize the transcriptomic signatures of tumor-infiltrating TREG cells, transitioning (4-1BB+) TREG cells and TFR cells. Elucidating the enhancer profiles in different developmental stages of TREG to TFR differentiation is likely to provide crucial insights into the TFs that instruct this differentiation step. These analyses will define genes and TFs that are pivotal for the heightened suppressive capacity of TFR cells, as well as for their maintenance or differentiation. Strategies to deplete TREG cells or to curb their functionality with the aim of enhancing anti-tumor immunity are being intensively investigated. Crucially however, most of these approaches are based on antibodies (i.e., ADCC-optimized for TREG cell depletion; i.e. anti-CTLA-4), which have an inherently long half-life in vivo, thus increasing the likelihood of causing irAEs. Conversely, our data imply that alternative dosing regimens of Phosphoinositide 3-kinase δ (PI3Kδ) inhibitors might offer a pathway to target TFR cells more specifically. We propose that PI3Kδ represents a novel and appealing immunotherapy target in solid tumors. In Aim 2 and Aim 3, we will assess whether alternative dosing regimens of PI3Kd inhibitors can be utilized to effectively and safely exploit the immunomodulatory impact of PI3Kδ inhibitors in solid cancers and whether a transient depletion or inhibition of TFR cells might suffice to restrict the immunosuppressive milieu in the tumor and thus drive anti-tumor immunity without causing toxicity.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY/ABSTRACT Although immune checkpoint inhibitors (ICIs) are revolutionizing cancer treatment, they are associated with life- or organ-threatening complications, termed immune-related adverse events (irAEs). Steroids, the first-line of treatment for irAEs, significantly abrogate the anti-tumor efficacy of ICIs; however, our knowledge of the mechanisms and signs that underpin the onset and progression of irAEs is very limited. Furthermore, commonality and individuality of altered immunity between irAEs have never been characterized. Our clinical and translational studies have revealed significant impact of combined ICI therapy (cytotoxic T lymphocyte- associated antigen 4 (CTLA-4) and programmed cell death protein 1 (PD-1) inhibitors) on the severity of arthritis-irAE, -colitis-irAE, and -pneumonitis-irAE compared to PD-1 inhibitor monotherapy. Notably, T helper (Th)17 cell signatures were significantly enhanced in arthritis-irAE synovial fluid, colitis-irAE colon, and pneumonitis-irAE bronchoalveolar lavage. In parallel, our lab has developed arthritis-irAE, colitis-irAE and pneumonitis-irAE murine models recapitulating patients’ clinical settings. Importantly, like in humans, Th17 cell signatures were enriched in the inflamed tissue of mice with ICI-induced arthritis (synovium), colitis (colon), and pneumonitis (lung) after combined ICI-therapy and correlated with irAE disease severity. Interestingly, arthritis-prone mice developed arthritis after receiving fecal microbial transplant (FMT) from combined ICI arthritis donor mice, notably with enhanced Th17 cell signatures. We also observed that blockade of Th17- related cytokines, interleukin (IL)-6 and tumor necrosis factor alpha (TNFα), may pinpoint irAE immunity while preserving/enhancing the anti-tumor efficacy of ICI therapy in humans and mice. Further understanding of mechanisms underlying irAEs may lead to identification of common and valid biomarkers predicting development and/or reflecting severity of irAEs as well as to new steps in their treatment. Therefore, we propose to uncover shared and distinct cellular/molecular mechanism(s) underpinning irAEs (arthritis- irAE, colitis-irAE, and pneumonitis-irAE) development as well as establish efficient therapeutic strategies. Here, in Aim 1, we will identify the cellular and molecular mechanisms and therapeutic targets by utilizing our novel preclinical murine models of irAEs including arthritis, colitis, and pneumonitis. In addition, cellular, phenotypic, and transcriptomic analysis of biospecimens from cancer patients with arthritis-irAE, colitis-irAE or pneumonitis-irAE will help to uncover common or unique immunopathogenesis mechanisms between different irAEs. In Aim 2, we will develop optimal strategies for treatment of irAEs while preserving anti-tumor immunity by utilizing melanoma tumor model in mice with irAEs. The implications from this work will be significant and will help to: 1) discover mechanisms universally or distinctively present in irAEs; 2) identify mechanistic biomarkers reflecting irAE disease activity; and most importantly, 3) develop a mechanism-rooted therapeutic strategy for irAEs without sacrificing anti-tumor immunity.

NIH Research Projects · FY 2025 · 2023-07

ABSTRACT Treatment options for small cell lung cancer (SCLC) patients have remained largely unchanged for 3 decades, with no new FDA-approved treatments for 20 years, and no targeted therapies. We recently performed a screen for targets of an arginine methyltransferase called CARM1 and identified the NFI family of transcription factors as substrates for this PRMT. Importantly, NFIB harbors both oncogenic and metastatic promoting activities in the context of SCLC development. We confirmed that CARM1 functions as a transcriptional coactivator for NFIB. Based on these finding, we hypothesize that CARM1 methylation of NFIB is critical for its tumor-promoting functions. To further support this premise, we have generated a Nfib knockin mouse that harbors a R-to-K mutation in the CARM1 methylation site. When this mouse is crossed onto a SCLC genetically engineered mouse model (GEMM) the life expectancy of these mice is lengthened by a third (from 200 to 300 days), which is almost identical to the impact of Carm1-loss in the same GEMM. These finding raise the possibility of targeting SCLC with CARM1 small molecule inhibitors. We have also identified an effector molecule (TRIM29) for the CARM1 methylation site on NFIB. In this proposal we plan to: (1) perform a deep mechanistic analysis of this newly discovered CARM1/NFIB/TRIM29 signaling axis, and (2) investigate the therapeutic potential of targeting this axis using a battery of pre-clinical mouse models.

NIH Research Projects · FY 2025 · 2023-07

ABSTRACT Immune checkpoint inhibitors (ICIs) have improved outcomes in metastatic non-small cell lung cancer (NSCLC), and providers may now choose between multiple first-line ICI-based regimens including ICI monotherapy and ICI with chemotherapy. However, this increase in options has complicated clinical management, with few biomarkers to guide upfront ICI treatment selection, and incomplete metrics for early on-treatment assessment of response to ICI therapy. Hence, there is an urgent need for novel analytics tools to optimize and personalize immunotherapy treatment strategies. While prior biomarker efforts have focused largely on tissue-based molecular profiling, these have demonstrated limited predictive power and are difficult to implement due to practical limitations in acquiring pre- and on-treatment tissue. In contrast, imaging and blood-based assays offer a unique and non-invasive mechanism by which the biology of the tumor and the changes on treatment can be studied and modeled. Thus, we propose an integrated radiomic-blood analysis to develop predictors of pre- and on-treatment response to guide the clinical management of NSCLC. Our primary goal is to develop radiomic- blood signatures for precision immunotherapy in advanced NSCLC by leveraging our expertise in data science, thoracic oncology, cancer genomics, computational oncology, clinical assay development, and established research collaborations. Our preliminary data demonstrates our success in utilizing multi-parametric profiling of circulating tumor DNA to identify molecular phenotypes associated with ICI outcome and disease recurrence, and in developing novel radiomic subtyping techniques with superior outcome prediction and demonstrated association with underlying lung cancer biology. Hence, we hypothesize that coupling radiomic and blood-based metrics can non-invasively inform therapeutic decision-making in NSCLC management while advancing our understanding of NSCLC biology. To advance this hypothesis, we have assembled a unique set of cohorts of metastatic NSCLC patients treated with ICI regimens with high-quality radiographic scans, blood samples, and molecular and clinical data: our in-house lung cancer database (GEMINI, n=5000); a validation dataset from our collaboration with the Massachusetts General Hospital (MGH) Cancer Center (MGH, n=600); the multicenter collaborative Stand Up 2 Cancer/Mark Foundation cohort (SU2C, n=400), and a prospective phase III ICI trial (LONESTAR, n=300). Our proposal builds on these unique cohorts and our promising preliminary data to construct predictive models to guide up-front ICI therapy selection and improve on-treatment response assessment, while complementary investigations will uncover the biology underlying these clinical predictors. A major strength of our proposal is our interdisciplinary team’s expertise in developing, validating, and translating these innovative predictive models toward highly relevant clinical questions. The development of integrative blood- and imaging-based radio-genomic biomarkers will help improve the clinical management of patients with metastatic NSCLC while helping progress the field toward a new era of non-invasive precision immunooncology.

NIH Research Projects · FY 2026 · 2023-06

For many reproductive-age cancer survivors, access to safe and effective methods for fertility preservation is an essential but often elusive path to creating a family. As more women delay childbirth and greater numbers are diagnosed before completing childbearing, fertility concerns have become increasingly relevant. Although the appropriateness of fertility treatment varies by clinical and patient factors, cancer survivors often require assisted reproductive technologies (ARTs) at younger ages and may face reduced chances of conception. Fertility-related needs arise in two key clinical contexts: at cancer diagnosis (for fertility preservation) and after treatment completion (to achieve pregnancy). In both situations, multiple factors—such as insurance coverage, geographic access to ART clinics, and neighborhood socioeconomic conditions—can affect access to services. Many patients pay out of pocket for ARTs, making this option unaffordable for those with limited financial means. Our study addresses two primary questions: 1. What are the sources and extent of variation in fertility preservation, ART use, and live births among cancer survivors? 2. Can expanded access to ARTs mitigate these variations in outcomes? Using a multilevel framework, we will examine how individual-, neighborhood-, and policy-level factors (e.g., state insurance mandates) influence ART access and use. Our specific aims are to: - Investigate factors associated with fertility preservation - Investigate factors associated with ART use after treatment - Assess how these factors contribute to live birth rates following cancer diagnosis Findings will clarify the barriers that affect family-building opportunities for reproductive-age cancer survivors and support evidence-based policy and service delivery improvements.

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY The maintenance of genomic integrity after DNA damage and replication stress depends on the coordination of DNA repair and cell cycle checkpoints. The replication checkpoint pathway, which comprises two critical protein kinases, ATR and CHK1, has an essential role in this coordination. Inhibition of the replication checkpoint results in cell lethality in response to replication stress induced by oncogenes, radiation, or chemotherapeutic agents. Indeed, several replication checkpoint inhibitors are being tested in clinical trials as potential anticancer agents. Our objective in this application is to provide a detailed mechanistic understanding of replication checkpoint control, which may help us develop the best strategies for using these checkpoint inhibitors in cancer therapy. We have studied replication checkpoint control for many years. We demonstrated that the ATR-dependent replication checkpoint can be activated by a variety of DNA lesions. In addition, ATR not only activates CHK1 but also phosphorylates many other substrates, including MCM, H2AX, and others at or near stalled replication forks to initiate replication checkpoint signaling. Moreover, we have identified and studied several key proteins such as TOPBP1, ETAA1, and Claspin involved in replication checkpoint control. Many key proteins involved in DNA replication and replication checkpoint control are essential for cell survival. Thus, it remains challenging to fully understand the roles of DNA replication and replication checkpoint proteins in cell cycle progression. Toward this end, we recently took advantage of the dTAG- mediated protein degradation system and established cell lines with inducible degradation of several essential DNA replication and replication checkpoint proteins. Initial analyses of events following the depletion of these essential proteins revealed some unexpected observations, which led us to re-evaluate replication checkpoint control and the mechanisms underlying cell cycle transitions. In this project, we will determine the essential functions of these DNA replication and checkpoint proteins in S phase and cell cycle progression. We anticipate that results from these studies will provide a better understanding of replication checkpoint control, especially how DNA replication and replication checkpoint are coordinated to ensure S phase progression and cell survival.

NIH Research Projects · FY 2026 · 2023-06

The Center for Transformative Community-Driven Research to Prevent Obesity-related Cancer (the Center) seeks to reduce cancer risk and ultimately improve cancer outcomes for communities. We plan to bridge the gap between research and real-world practice through community-engaged approaches to address obesity—the second leading preventable cause of cancer. The Center will focus its activity on Acres Homes, a community in Houston, Texas that has high prevalence of cancer risk factors. The Center will test interventions in the context of MD Anderson’s current place-based cancer prevention initiative (Be Well Communities™), around two scientific themes: 1) understanding social and environmental context in which risk factors arise (e.g., food insecurity, limited recreational infrastructure, few health promotion services) and 2) promoting health behavior changes of diet and physical activity as major contributing causes of obesity. The two research projects span the cancer control continuum from primary prevention to survivorship: Project 1 will test the effect of a multi-level nutrition intervention on BMI and metabolic health in Acres Homes elementary school students and their parents, and Project 2 will adapt and test the effect of a physical activity intervention for Acres Homes cancer survivors on physical functioning. Both will use a health promotion interweaving approach to embed the interventions in the environmental context of Acres Homes and Be Well Communities. The interdisciplinary approach used by the research projects will integrate behavioral science, implementation science, geospatial analytics, and health communication. The Center will include five cores: Administrative, Research and Methods, Career Enhancement, Developmental, and Engagement Core. Our conceptual framework is informed by both the environmental context and the science of behavior change. At the behavior change level, the conceptual framework incorporates the COM-B (Capability, Opportunity, Motivation -Behavior) model of behavior change. In collaboration with Acres Homes community leaders, the specific aims of the Center are to: (1) Test interventions to address risk factors for obesity-related cancers using a health promotion interweaving approach in the context of Be Well™ Acres Homes, building on the existing infrastructure of community collaboration, planning and programming; (2) Collaborate with residents and community organizations to foster an environment that supports the conduct of research and the implementation of programs that focus on exploring risk factors for obesity-related cancers; (3) Support the careers of interdisciplinary teams of early-career researchers including trainees and early stage faculty and build their expertise in conducting research to promote cancer prevention and control.

NIH Research Projects · FY 2026 · 2023-06

One strategy to enhance the immune response to tumors is radiotherapy (RT). Recent data support that RT- induced micronuclei (MN) are intrinsically immunostimulatory, as ruptured MN releases double stranded DNA (dsDNA) eliciting the cycling GMP-AMP synthase (cGAS) and stimulator of interferon genes (STING) pathway. Although progress has been made in combining immune checkpoint blockade (ICB) with RT, relatively little is known about the physical mechanisms of RT that elicit immunostimulatory signals and how they can be har- nessed clinically in the context of DNA damage and DNA repair inhibition. Radiation with high ionization density (or linear energy transfer, LET) induces more clustered DNA lesions, more MN and higher cell kill compared to low-LET radiation. α-particles are characterized by their high-LET in contrast to photons and protons and may be ideal for creating high levels of MN and downstream enhanced activation of immunostimulatory signals through the cGAS-STING pathway. A novel modality to deliver α-particles has recently been successfully demon- strated in a phase I clinical trial using a method called diffusing alpha-emitters radiation therapy (DaRT). DaRT consists of interstitial radioactive seeds coated with radium-224, an α-particle emitter. The radium-224 decay chain is unique in that the decay products also emit α-particles and diffuse, allowing the α-particles’ dose to be deposited over 2-3 mm from the seed. Thus, multiple seeds implanted into a tumor allow the high-LET α-particle dose to be deposited within the entire tumor volume. In addition to radiation, pharmacologic inhibition of DNA repair affects the presence of MN. This inhibition of DNA repair can be created with drugs such as Ataxia telan- giectasia and Rad3 related (ATR) inhibitors (ATRi). The combination of an ATRi with α-particle-induced clustered DSB lesions may synergistically enhance the accumulation of MN, ultimately enhancing immunostimulatory sig- nals. These will be investigated with ICB to determine how to synergistically augment RT-induced antitumor immunity. We hypothesize that α-particles combined with an ATRi produces more MN, results in more cGAS binding to dsDNA and consequently potentiate robust antitumor immunity. We propose to: 1) Elucidate the mech- anisms by which cGAS binds to dsDNA in -particles+ATRi treated cells; 2) Elucidate the mechanisms by which -particles+ATRi induces immune signaling; and 3) Evaluate antitumor immunity from -particles+ATRi in vivo. Our research has the potential to define -particles as a tool to augment antitumor immune response, especially for tumors that are known to be unresponsive to ICB. Our proposed research is of critical relevance to address the poor prognosis associated with multiple advanced solid cancers that are immunologically cold. Our proposed work is innovative, in that it aims to define the effects of -particle-induced clustered DNA damage on tumors in the context of antitumor immunity. Our findings will elucidate the mechanisms behind immune modulation by high-LET radiation, which may ultimately guide the combined rational use of modalities that use high-LET radi- ation (including α-emitting radiopharmaceuticals), DNA repair inhibitors and ICB for aggressive cancers.

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY/ABSTRACT Myeloid cell-mediated immune suppression is one of the major factors responsible for resistance to anti- programmed cell death protein 1 (PD-1) therapy. Glioblastoma (GBM), a brain tumor with a dismal prognosis with current standard-of-care therapy, is enriched with immune-suppressive myeloid cell subsets in its tumor microenvironment and shows resistance to anti-PD-1 therapy. Therefore, there is an unmet need to develop strategies to overcome myeloid-derived immune suppression in order to provide durable clinical benefits of anti- PD-1 therapy in patients with GBM. Epigenetic machinery plays a key role in myeloid cell differentiation and establishing specific functional profiles. However, the impact of epigenetic regulation of intra-tumoral myeloid cells on resistance to immunotherapy has remained unexplored. The overall objective of the current proposal is to identify key epigenetic factors regulating immune-suppressive pathways and develop a novel strategy targeting the epigenetic regulator(s) to reverse myeloid-derived immune suppression in GBM. In our preliminary studies, we noted that immune-suppressive myeloid cell subsets in human GBM tumors have high expression of an epigenetic enzyme - histone 3 lysine 27 demethylase (KDM6B). GBM tumor-bearing mice carrying myeloid-cell specific Kdm6b deletion demonstrated improved survival. Additionally, the absence of Kdm6b increased chromatin accessibility and expression of genes associated with proinflammatory pathways including interferon response, phagocytic ability, and antigen-presentation in intratumoral macrophages and dendritic cells. Importantly, pharmacological inhibition of KDM6B with GSK-J4, a small molecule inhibitor, enhanced the efficacy of anti-PD-1 therapy in GL261 tumor-bearing mice with increased infiltration of effector T cells. Based on the preliminary findings, we hypothesize that KDM6B inhibition reprograms immune- suppressive myeloid cells into a proinflammatory phenotype, thereby enhancing T cell-mediated anti-tumor immunity to overcome resistance to anti-PD-1 therapy in GBM. In the current proposal, we will test our hypothesis using three specific aims: 1) To determine the mechanism of KDM6B-mediated functional and epigenetic regulation of phagocytosis and antigen presentation; 2) To identify the role of KDM6B in myeloid cell-mediated regulation of T cell function and localization; and 3) To evaluate the therapeutic potential of KDM6B inhibition in reversing the resistance to anti-PD-1 therapy. The research is innovative in the applicant’s opinion because this proposal will be one of the first to provide systems-level understanding of the role of epigenetic regulation of myeloid cell biology at a single-cell resolution in GBM. The proposed research is significant since it will investigate a novel strategy of targeting the epigenetic machinery to overcome myeloid cell-derived immune resistance to anti-PD-1 therapy in GBM. The long-term goal of this research endeavor is to develop personalized immunotherapies with epigenetic modulators in a tumor-specific manner.

NIH Research Projects · FY 2026 · 2023-06

ABSTRACT Our overarching goal is to evaluate the therapeutic potential and mechanism-of-action of the epigenetic regulatory factor NSD2 in lung adenocarcinoma (LUAD). Lung cancer is the most common cause of cancer- related mortality in the United States and worldwide, leading to over a 1.8 million deaths each year. LUAD is its most common histological type of lung cancer. While new targeted and immunotherapies have improved median survival for LUAD patients, unfortunately, improvement in outcomes over the past 20 years has been incremental. Thus, there is great interest in identifying novel factors that might cooperate with the canonical oncogenic pathways that drive LUAD with the notion that a therapeutic strategy hitting multiple pathways will mitigate potential drug toxicity by lowering the overall dose needed for each medicine and in parallel combat resistance development. A central hypothesis to be tested here is that the clinically actionable and histone lysine 36 (H3K36) di-methyltransferase enzyme NSD2 is such a factor. In preliminary work we found that NSD2 promotes aggressive malignant tumor progression and rapid lethality in a classis LUAD mouse model. In our proposal, we will investigate the molecular, epigenetic, cellular, and in vivo role of NSD2-H3K36me2 axis in lung cancer and directly test the efficacy of NSD2-targeted therapy using a first-in-class NSD2 inhibitor and state-of- the-art pre-clinical models of LUAD. In Aim 1 we investigate the role of NSD2 in lung cancer pathogenesis. We have developed a KRASG12C- driven NSD2 tunable mouse lung cancer model to investigate the function and mechanism of action of elevated or depleted NSD2 activity in LUAD initiation, progression, metastasis, and intratumoral heterogeneity. In Aim 2 we evaluate therapeutic efficacy of NSD2 inhibition using a first-in-class potent inhibitor of NSD2. We will utilize genetic models such as LUAD driven by KRASG12C mutations and multiple patient derived LUAD xenografts to test anti-tumor efficacy of NSD2 inhibition as a single agent and as part of combination therapies. Together, this work will be the first to evaluate the therapeutic potential and mechanism-of-action of NSD2 in LUAD.

NIH Research Projects · FY 2026 · 2023-06

Project Summary We demonstrated that short-term fasting protects mice from high dose ionizing radiation by protecting small intestinal (SI) stem cells, thereby preserving SI homeostasis and promoting organismal survival. We also showed that the major ketone body produced by fasting directly modifies the epigenome of SI epithelial cells to induce gene expression changes and that fasting alters the gut microbiome to favor bacteria whose metabolites are known to induce epigenetic and gene expression changes. We will test the hypothesis that ketone bodies and microbial metabolites induced by fasting, cause epigenetic and gene expression changes in SI epithelial cells thereby providing GI radioprotection.

NIH Research Projects · FY 2026 · 2023-05

Project Summary Current approaches to treating triple-negative breast cancer (TNBC) remain unsatisfactory. There remains an unmet need to establish novel alternative therapies that can both exert targeted effects on cancer cells as well as stimulate an anti-cancer immune response that may render additional opportunity for combination with emerging immunotherapy. We have uncovered a novel onco-metabolic feature in TNBC wherein cancer cells exploit a sphingomyelin lipid scavenging phenotype that is targetable through repurposing of the selective glucosylceramide synthase inhibitor eliglustat, an FDA approved drug for treatment of Gaucher Disease. Our preliminary studies demonstrate that eliglustat promotes accumulation of mitotoxic ceramides with resultant shift from survival mitophagy to lethal mitophagy and subsequent cancer cell death. Moreover, we show that eliglustat suppresses tumor growth at clinically achievable doses in TNBC tumor-bearing mice and that the anti-cancer effects of eliglustat are associated with pronounced increases in tumor infiltrating CD4+ and CD8+ T-cells, suggesting an additional role of eliglustat in potentiating an anti-tumor immune response. The primary objectives of this proposal are to establish novel utility for eliglustat as an ‘immunometabolic adjuvant’ for the treatment of TNBC and to also define the mechanism(s) by which eliglustat potentiates an anti-cancer immune response. To test this, we will assess the anti-cancer efficacy of eliglustat in patient-derived xenograft (PDX) models generated from patients with primary treatment-naïve TNBC that did or did not go on to respond to chemotherapy (Specific Aim 1a). To test whether the combination of eliglustat plus anti-PD-L1 yields improved anti-cancer effects compared to either treatment alone, we will use the BRCA1co/co; MMTV-Cre; p53+/- and 4T1 orthotopic syngeneic mouse models of TNBC (Specific Aim 1b). Primary endpoints of interest for in vivo studies will be overall survival and tumor growth; effects on the tumor immunophenotype following intervention will be assess using multiplex immunofluorescence panels and single cell transcriptomics for single cell-level expression profiling of tissues. We also aim to define the mechanisms by which eliglustat induces an anti-cancer immune response. First, we will evaluate the effect of eliglustat on cGAS-STING signaling proteins and downstream pathway activities in TNBC cells (Specific Aim 2a). Next, we will use advanced mass spectrometry technologies coupled with novel isolation methods for extracellular vesicles (EVs) to define the MHC-I bound peptidome on surfaces of TNBC cells and TNBC-derived circulating and intra-tumor EVs following eliglustat treatment. ELIspot and Cytotoxic T Cell-based tumor killing live-cell assays will be used to test the functionality of EVs on activating T-cells ex vivo (Specific Aim 2b). If successful, our potential findings will provide key pre- clinical evidence for the use of eliglustat and justification for moving eliglustat into early phase clinical trials. Given that eliglustat is already FDA-approved for another indication, it has high potential to be readily translated into clinical use, and potentially providing a new paradigm for cancer immunotherapy.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY/ABSTRACT Oncogenic KRASG12C (KG12C) mutations underpin the development of ~14% of non-squamous non-small cell lung cancer (NSCLC) and account for ~10,000 deaths annually in the U.S. The development of potent, selective and clinically active covalent inhibitors of the KG12C oncoprotein represents one of the most exciting recent advances in the field of targeted cancer therapy, yet strategies to circumvent the development of adaptive resistance and improve the durability of individual responses to KG12C inhibitors are urgently needed in order to transform clinical outcomes for patients. The role of crosstalk between tumor cells and the tumor microenvironment (TME) in the development of acquired KG12C inhibitor resistance in NSCLC has not been systematically examined to date, despite evidence that in more than 50% of cases no genomic resistance mechanisms can be identified at the time of radiological progression. Furthermore, key mediators of TME remodeling and immune escape in response to KG12C inhibitor therapy remain poorly defined and therapeutic strategies that target the adverse TME in order to prevent, delay or overcome KG12C inhibitor adaptation/acquired resistance have not been established. Finally, the impact of major co-occurring genomic alterations in STK11/LKB1 and TP53 and that shape the immune contexture of KRAS-mutant NSCLC– on non-genetic mechanisms of acquired KG12C inhibitor resistance is not known. Based on our preliminary findings and previous work we hypothesize that: 1. Remodeling of the tumor microenvironment and immune escape can promote non-tumor cell autonomous adaptation/acquired resistance to KG12C inhibitors; 2. Targeting STAT3 signaling with TTI-101 can forestall and possibly overcome non-genetic acquired resistance to KG12C inhibitors through effects on tumor cells and/or the TME. In Aim 1, we will determine the contribution of TME remodeling and immune escape to acquired KG12C inhibitor resistance in NSCLC, using immune competent models of KG12C NSCLC that recapitulate its co-mutational complexity. We will further interrogate the role of master mediators of TME adaptive remodeling with initial focus on STAT3 and we will validate key findings using paired biopsies from patients with metastatic KG12C-mutant NSCLC that were treated with sotorasib as part of standard of care. In Aim 2, we will evaluate the anti-tumor efficacy and TME- modifying effects of STAT3 inhibition with TTI-101 in combination with direct KG12C inhibitors in immune- competent models of KG12C NSCLC. Clinical significance: This work will yield fresh insights into non-genetic mechanisms of acquired resistance to KG12C inhibitors that rely on tumor-TME crosstalk and will facilitate the development of novel therapeutic strategies that tackle the adverse TME in order to maximize long-term clinical benefit from KG12C inhibitors.

NIH Research Projects · FY 2026 · 2023-05

Project Summary/Abstract The goal of this proposal is to develop statistical methods for evaluating treatment strategies at different time points and identifying optimal treatment strategies on the basis of patients' longitudinal biomarker measurements. It is motivated by our research on identifying the best timing for patients with chronic myeloid leukemia (CML) to receive a stem cell transplant (SCT). SCT can cure leukemia, but it is associated with life- threatening risks. For this reason, most patients start with other less-aggressive treatment options that are much safer but cannot cure the disease. Thus the decision-making about optimal timing of SCT depends on a patient's disease progression. However, it is infeasible to conduct a randomized controlled trial to weigh the risks and benefits of SCT at various times. To optimize this decision-making process, sophisticated and comprehensive statistical models are needed to provide an accurate estimation of the benefits and risks (and their trade-offs) over time for patients under different SCT timing options. However, these have not yet been developed, due to the challenges elaborated below. First, the question of an optimal decision on SCT cannot be answered by a single statistical model, it requires assembling information from a series of models and analyses. Second, there most likely is not a uniform solution for this question, as the optimal timing of SCT depends on each individual's disease progression status. Consequently, physicians must use patients' longitudinal biomarker trajectories to monitor their health status and make treatment decision in a dynamic fashion. Third, the treatment decision for each individual must account for their competing risks, including death by treatment-related complications and other causes (e.g., heart diseases and diabetes). Finally, it is impossible to implement optimal decision-making without an easy-to-use software. The following specific aims are proposed to solve these problems. Aim 1: Use functional component principal component analysis (FPCA) techniques to fully capture the dominant patterns from patients' longitudinal biomarker trajectories, and use them as predictors of patients’ risk of disease progression. Aim 2: Estimate dynamic competing risks based on baseline covariates and longitudinal biomarker trajectories using multi-state models. Aim 3: Use analytic and microsimulation approaches to estimate and compare the mean survival times under different SCT timing options. Aim 4: Conduct validation studies, develop software, and broaden application. Three CML studies will be used to cross-validate each other regarding the optimal timing of SCT. Software programs with user-friendly interfaces will be made publicly available. The proposed statistical and software programs will be adapted and applied to a study of kidney disease to test their broad application.