University Of Tx Md Anderson Can Ctr

NIH Research Projects · FY 2026 · 2023-05

Project Summary/Abstract Oncogenic mutations in KRAS represent the most prevalent genomic driver event in lung adenocarcinoma (LUAD) (~30%) and account for ~25,000 deaths annually in the U.S. Immunotherapy (IO) with immune checkpoint inhibitors (ICI) is active in KRAS-mutant non-small-cell lung cancer (NSCLC), however only a minority of patients exhibit long-lasting responses. Co-occurring genomic alterations can shape the immunobiology of KRAS-mutant NSCLC and impact its response to ICI. Loss-of-function somatic mutations in RBM10, encoding a regulator of alternative splicing (AS), are prevalent in LUAD (~8%) and are significantly enriched in KRAS- mutant NSCLC (~25%). We found that loss of RBM10 in KRAS-mutant NSCLC tumors and cell lines results in DNA double-strand breaks (DSBs) and STING-dependent but cGAS-independent NF-κB signaling, that establish an immunosuppressive tumor microenvironment (TME) – rich in MDSCs and M2-macrophages – and support tumor immune escape. Critically, RD NSCLC exhibit selective sensitivity to anti-CSF1R, that depends on functional CD8+ T-cells. Preliminary evidence points to accumulation of R-loops and distinct STING isoforms as candidate mechanisms that underpin DDR activation and preferential NF-κB engagement in RD cells. Based on our preliminary findings we hypothesize that: 1. In KRAS-mutant NSCLC, RBM10 loss triggers R-loop accumulation and aberrant DDR signaling that support STING-dependent but cGAS-independent pro- tumorigenic NF-κB signaling; 2. Splicing dysregulation upon RBM10 loss promotes STING isoforms that preferentially engage NF-κB over TBK1 and IRF3; 3. RBM10 loss remodels the NSCLC TME and fosters immune evasion 4. The sensitivity of RD NSCLC to ICI can be enhanced by co-targeting STAT3 with TTI-101. In Aim 1, we will dissect the link between RBM10 loss, DDR activation and STING-mediated NFκB signaling and we will assess the contribution of altered R-loop homeostasis and alternative STING splicing to these phenotypes. In Aim 2, we will comprehensively characterize the composition, signaling pathways and functional properties of the RD TME in preclinical models in order to identify critical mediators of immune evasion and we will validate key findings in NSCLC clinical specimens. Finally, in Aim 3, we will determine the impact of RBM10 inactivation on the clinical efficacy of ICI using clinical outcome data/specimens from patients enrolled in two phase 3 clinical trials of durvalumab with or without tremelimumab versus platinum-doublet chemotherapy for previously untreated metastatic NSCLC as well a phase 3 clinical trial of nivolumab/ipilimumab. In addition, we will evaluate co-targeting STAT3 in combination with anti-PD-(L)1 in order to enhance the efficacy of immune checkpoint blockade in RD-NSCLC. Clinical significance: This work will examine a novel link between splicing dysregulation and immune evasion that is mediated by STING and will further seek to develop precision combination immunotherapy approaches for RD NSCLC. The strength of our assembled multi-disciplinary team of experts will facilitate rapid translation of discoveries into clinical advances for NSCLC patients.

NIH Research Projects · FY 2026 · 2023-05

Breast cancer accounts for one in four cancer diagnoses in women, affecting up to one in eight women in the United States. All patients with breast cancer undergo surgery, yet surgery is not without risks and is associated with a multitude of side effects. Many of the complications from surgery are from the use of general anesthesia and intra- and postoperative opioids. Side effects of general anesthesia include hemodynamic instability, postoperative nausea and vomiting, suppression of cell-mediated immunity, cognitive impairment, and delayed recovery. The use of opioids is also associated with postoperative nausea and vomiting, ileus, urinary retention, pruritus, and immunosuppression. Frequently, intra- and postoperative opioid use often leads to an increased risk for non-medical opioid use and opioid-sparing techniques are needed. Extensive data supports the use of non-pharmacological interventions including hypnosedation (HS) for patients undergoing invasive medical procedures. In a variety of medical populations, patients using HS report significantly less anxiety and pain, demonstrate beneficial physiological responses, request less analgesic medication, and spend less time in the procedure room than controls. In a number of studies, the improved patient-reported outcomes and overall satisfaction was also paralleled by decreased medical costs. All of these previous studies either delivered the HS during simple medical procedures (e.g., breast biopsy) or before more invasive procedures such as breast surgery and no RCTs have provided hypnosis during surgery delivered by one of the surgical team members. The proposed trial will randomize women and men with breast cancer scheduled for a lumpectomy ± sentinel node biopsy to one of three groups: 1) surgery with a local anesthetic, fentanyl, and HS before and during surgery (HS); 2) HS before surgery with usual care general anesthesia (HS-GA; propofol infusion, fentanyl, and local anesthetic); or 3) Usual care general anesthesia (GA). The study will examine differences in opioid use, pain, anxiety, nausea, fatigue, and cognitive dysfunction before and after surgery. Recovery will be also tracked 14 and 90 days after surgery. This project will allow further exploration of HS during surgery provided by a clinical team member and to explore the biopsychosocial processes associated with analgesia and opioid use, anesthesia, and pain and examine baseline individual difference factors associated with the intervention effects and recovery. We specifically propose to test the hypothesis that HS during breast cancer surgery will result in better analgesia control along with lower opioid use, less pain and psychological stress, and faster recovery than GA. Data from the proposed study will help to move this intervention into the standard of care and expand the use of HS into other invasive medical procedures where general anesthesia may be avoided.

NIH Research Projects · FY 2026 · 2023-04

Research Summary The functional roles and molecular mechanisms of non-coding RNAs (lncRNA) in human genetic disease serve as promising new frontiers for establishing the foundation of future research directions and therapeutic avenues for disorders that currently lack sufficient treatment options. The inborn metabolic disorder phenylketonuria (PKU) has been considered to be caused by mutations in the phenylalanine hydroxylase (PAH) gene, resulting in disruptions to its enzymatic activity and consequent accumulation of phenylalanine in the blood. Our preliminary data demonstrated that the human lncRNA HULC is functionally conserved with mouse lncRNA Pair. PKU patients harbor genomic variants of the HULC gene. Knockout of Pair leads to hypo-pigmentation, growth retardation, progressive neurological symptoms, and seizures, which faithfully models human PKU. Deletion of HULC in primary human hepatocytes leads to elevated cellular Phenylalanine, suggesting the functional importance of HULC/Pair in PKU. We defined the RNA motifs of both HULC and Pair that are integral to establishing lncRNA-enzyme interactions and designed peptide-tagged lncRNA mimics that restored the enzymatic activity of PAH. Administration of HULC mimics in PKU mouse model successfully alleviated excess phenylalanine levels in serum, providing mechanistic insight into the basis of inherited genetic diseases. The proposed study seeks to establish long non-coding RNAs as important players in the initiation and progression of inborn metabolic disorders and develop a lncRNA-based innovative approach for determining new strategies to tackle the molecular basis of PKU. The central hypothesis is that the genetic mutation and deletion of lncRNA results in the enzymatic deficiency of PAH, which could be alleviated through targeting therapies to enhance the protein stability and enzymatic activity of PAH. Using iPS-differentiated hepatocytes derived from PKU patients, we will first demonstrate the biological significance of HULC in the development and progression of PKU by addressing the impact of common HULC mutations and assessing the correlations between mutations or deletions of HULC and PKU symptoms. We will then elucidate the molecular mechanisms of lncRNA-mediated regulation of PAH enzymatic activity. Finally, we will use the administration of liver-enriched lncRNA mimics and small molecule agonists/antagonists to establish their mechanistic role in restoring impaired enzymatic function. The aforementioned research aims will not only elucidate the roles of lncRNAs in metabolic disorders, but also illustrate the potential therapeutic value of lncRNA mimics and small molecule inhibitor-based medicine in treating inborn genetic diseases. We anticipate that the results of this study will shift the current research and clinical paradigm to incorporate the physiological and pathological aspects of non-coding genes and pave the way for the development of future lncRNA-based medicines.

NIH Research Projects · FY 2026 · 2023-04

Summary: CD19 directed CAR-T cells have transformed the treatment landscape of B-cell lymphoid malignancies. However, despite high initial complete remission rates, relapses occur within the first year of therapy in approximately 50% of patients who receive commercially available autologous CAR19 T-cells. Relapses can be classified into two patterns: CD19-positive relapse related to CAR T-cell exhaustion and senescence, or CD19- negative relapse related to target antigen loss. Patients who relapse after CAR19 T-cell therapy have poor prognosis; hence, there is an urgent need to develop the next-generation of CAR engineered immune effector cells that target tumors with efficacy and with minimal toxicity. There is growing interest in natural killer (NK) cells as a candidate for CAR therapy as they may prevent antigen escape through their innate ability to kill tumor cells and they are safe and well-suited for use in the allogeneic therapy setting. In a first-in-human study, our group showed the safety and promising activity of cord blood (CB) derived CAR-NK cells targeting CD19 in patients with B-lymphoid malignancies (Liu et al NEJM 2020). This proposal aims to build on this platform to develop the next-generation NK cell therapies by enhancing NK cell potency and persistence through optimal costimulatory signaling, cytokine armoring and checkpoint inhibition. We have identified CD70, as a novel therapeutic target in patients with B-NHL after CAR19 T-cell failure and developed a novel strategy to target CD70 by genetically modifying CB-NK cells with a retroviral vector (iC9-CD27-DAP10-CD3ζ-IL-15) that incorporates (i) the gene for a truncated human CD27 (the natural receptor for CD70) to redirect their specificity; (ii) DAP10 as an NK-specific costimulatory domain linked to a CD3ζ signaling endodomain; (iii) IL-15 to support their survival and proliferation, and (iv) inducible caspase-9 (iC9) as a suicide gene. Our preliminary data show the efficacy and safety of this approach in vitro and in vivo and support its translation to the clinic. In addition, we have developed a robust strategy to cryopreserve CAR-NK cells, allowing for the generation of a biobank of off-the-shelf engineered NK cells, thus reducing cost and increasing accessibility. Finally, we have devised a novel strategy to target the immune checkpoint CIS in our CAR-NK cells to modulate their metabolic fitness and potency. We hypothesize that targeting CD70 with iC9/CAR27D10ζ/IL-15 NK cells will greatly improve outcomes in patients with NHL after CAR19 T-cell failure and that by targeting the immune checkpoint CIS we can further enhance the CAR-NK cells’ metabolic fitness and potency. These concepts will be evaluated in three specific aims: In Aim 1 we will conduct a Phase I/II clinical trial to test the safety and efficacy of iC9/CAR27D10ζ/IL-15 NK cells in patients with CD70+ NHL who have failed CAR19 T-cell therapy (FDA approved, IND #27757). In Aim 2 we will apply innovative single-cell proteomic and transcriptomic studies to comprehensively characterize the fate of the CAR-NK cells and to identify key mechanisms of efficacy and resistance. In Aim 3, we will perform mechanistic studies to elucidate how CIS deletion enhances the metabolic fitness of CAR-NK cells and will perform IND enabling studies in preparation for the next-generation clinical studies testing CIS deficient iC9/CAR27D10ζ/IL-15 NK cells.

NIH Research Projects · FY 2026 · 2023-04

Project Summary This project, proposed by clinician scientist Stephen Chun, M.D. at the University of Texas, M.D. Anderson Cancer Center (MDACC) aims to expand the National Cancer Institute (NCI)-sponsored radiation oncology trial program in the Houston Area Location (HAL) integrated academic satellites and support his national leadership efforts with the NCI-sponsored cooperative groups. Since joining the MDACC faculty, Dr. Chun has led the establishment of the HAL radiation clinical trial program and is the Director of Radiation Oncology Clinical Research for the MDACC HALs. Under his leadership, there has been an unprecedented increase (1% in 2016 to 14% in 2021) in the proportion of HAL patients enrolled in radiation clinical trials. With the NCI-sponsored cooperative groups, Dr. Chun currently serves as national Principal Investigator (PI) of a high-profile NRG Oncology analysis, study champion of NRG/Alliance A082002, MDACC institutional PI of NRG-LU005, and has been a top enroller for NRG Oncology-RTOG 1308 that was previously in jeopardy for closure due to poor accrual. To achieve NCI-funded trial objectives in the MDACC HALs, Dr. Chun will spearhead the implementation and expansion of a translational biomarker program, neurocognitive testing capabilities for NCI-sponsored central nervous system (CNS) cancer trials, and the continuing medical education program on clinical research. Dr. Chun is passionate about establishing workflows to offer clinical trials to people of all backgrounds to reduce demographic disparities. In order to develop new research capabilities, Dr. Chun will join the leadership of the Protocol in a Day Workshop, which is responsible for developing MDACC investigator-initiated radiation trials, establish the radiation clinical research program at new and planned MDACC facilities, leverage artificial intelligence to capture more patients eligible for trials, and continue to develop concepts with the NCI-sponsored cooperative groups. He will commit a minimum of 20% of his effort (2.4 calendar months) to NCI-sponsored clinical research, and MDACC is committed to support Dr. Chun’s efforts and conduct of NCI-supported clinical trials. The primary objective of his career is to prioritize and expand institutional and NCI-sponsored trials in the MDACC HALs which aligns with this R50 funding mechanism as well as with the broader mission of NCI.

NIH Research Projects · FY 2026 · 2023-04

Mechanisms of Epigenetic Plasticity in Neuropathic Pain The major objective of our project is to determine how traumatic nerve injury impacts epigenetic regulatory networks involved in chronic pain. Neuropathic pain remains a major clinical problem and therapeutic challenge. Both sustained changes in gene expression in primary afferent neurons and synaptic plasticity at the spinal cord level are essential for to the development of chronic pain. α2δ-1 (encoded by the Cacna2d1 gene) is a clinically validated neuropathic pain target and mediates the therapeutic actions of gabapentinoids. Traumatic nerve injury and certain cancer chemotherapeutic drugs cause α2δ-1 upregulation in the dorsal root ganglion and spinal cord, which augments nociceptive input to spinal dorsal horn neurons by directly interacting with NMDA receptors. Yet we know almost nothing about how nerve injury initiates and sustains the high expression level of α2δ-1. Acetylation of lysine residues in histone tails is dynamically regulated by various histone deacetylases (HDACs). However, the specific HDAC subtypes responsible for the upregulation of α2δ- 1 and other neuroplasticity-related genes in neuropathic pain have not been rigorously studied or identified. To address this key knowledge gap, we will specifically determine the role of class I HDAC subtypes in the control of histone acetylation and expression of α2δ-1 and other gene targets implicated in synaptic plasticity in two neuropathic pain models. On the basis of our preliminary data, we propose to test the overall hypothesis that nerve injury and chemotherapy diminish HDAC2 occupancy to induce histone hyperacetylation, via CK2- mediated phosphorylation, at the promoters of Cacna2d1 and other neuroplasticity-related genes in the DRG and that HDAC2 constitutively restrains chronic pain by repressing Cacna2d1 transcription and α2δ-1– dependent NMDA receptor activation at the spinal cord level. We will apply several innovative and vigorous approaches, including unbiased genome-wide epigenetic analyses, transgenic mice, and synaptic recordings to study neuroplasticity at molecular, cellular, and behavioral levels. Our project will generate fundamental new information about the epigenetic basis of neuropathic pain. Findings from our project are expected to advance our knowledge of molecular mechanisms of epigenetic plasticity and to guide the development of new strategies for treating neuropathic pain.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Cellular turnover is essential for the form and function of epithelial tissues. The rate of cell turnover slows during aging, can be accelerated during injury and repair, and is precociously stimulated during carcinogenesis; but the mechanisms guiding it in living tissues is not well understood. We recently discovered that elimination of unfit cells by extrusion from epithelia stimulates proliferation of nearby stem cells to replace the lost cells and maintain overall cell numbers in the tissue. This intriguing finding suggests extrusion provides a key link between cell loss and proliferation, and thereby controls the rate of cell turnover. Thus, identification of the mechanisms that underlie extrusion may provide new insights into endogenous processes that can be leveraged to promote cellular replacement or prevent the unwanted addition of new cells. Our long-term goal is to define the cellular and molecular mechanisms underlying the rate of cellular turnover in epithelial tissues. Using the developing zebrafish to study cell extrusion in a living epithelial tissue, we have found that cells fated to extrude alter their mechanical properties in the form of pulsatile actomyosin contractions that are controlled by enrichment of the bioactive lipid sphingosine-1-phosphate (S1P). We have also interrogated the cell loss-induced signaling events and cellular responses, including inflammatory cell recruitment and epidermal cell proliferation, that drive turnover. We identified a significant upregulated expression of the epidermal growth factor receptor ligand epigen (EPGN) upon induced cell extrusion, suggesting that transient increases in EPGN may aid in sustaining epithelial form and function during cell loss. Consistent with this idea, we found that treatment with recombinant human EPGN (hrEPGN) suppressed epithelial cell extrusion after receiving damage stimuli, which in turn reduced the compensatory stem cell proliferation. These data led to the hypothesis that EPGN regulates extrusion to dictate the rate of cellular turnover in epithelial tissues. One formidable challenge to studying cellular turnover and testing this hypothesis in a living organism involves visualizing and perturbing the complex interplay between extruding cells, the surrounding stem cells that replace the lost cells and immune cells to sense and respond to disruptions in integrity. Therefore, we created tools to manipulate different cellular and molecular components individually or in combination in living epithelial tissues of developing zebrafish and analyze changes to turnover in the presence of an innate immune system. Our work over the next five years we utilize this new approach and will focus on three essential areas that emerged from our ongoing studies and address key gaps in our knowledge of cellular turnover. First, we will determine the mechanisms regulating the localized changes in physical forces that are required to remove defective cells by extrusion. Second, we will determine how cell loss promotes changes in the epigenetic and transcriptional states in surrounding stem cells to stimulate proliferation and replace the lost cells. Third, we will determine the role of the innate immune system in promoting cell turnover and maintenance of epithelial tissue homeostasis.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT Our overarching goal is to elucidate the mode of action of and evaluate the therapeutic potential of the epigenetic regulatory factor NSD3 in the regulation of lung squamous cell carcinoma (LUSC) pathogenesis. 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. LUSC is the second most common subtype of lung cancer, accounting for ~30% of all cases and tragically over 40,000 deaths each year in the US alone. While new targeted therapies have shown promise in other malignancies, unfortunately, to date, there are no approved targeted therapies for LUSC. Thus, there is a major unmet need to uncover new, clinically actionable, and compelling targets for the development of new medicines to ultimately treat this difficult disease. A central hypothesis to be tested here is that the histone H3 lysine 36 (H3K36) di-methyltransferase enzyme NSD3 is a promising epigenetic target for the treatment of LUSC. In preliminary work we found that NSD3, which is commonly amplified in LUSC, is a major driver of LUSC pathogenesis in mouse and human models of this cancer. In our proposal, we will investigate the role of the NSD3-H3K36me2 axis in lung cancer in vivo and explore the molecular and epigenetic basis of NSD3-driven tumorigenesis. In Aim 1 we investigate the role of NSD3 in LUSC pathogenesis. We have developed novel mouse models that recapitulate the most common genetic alterations in human LUSC, including NSD3 amplification, and incorporated an inducible dual-recombinase approach to allow study of multi-step tumorigenesis in vivo. This system will be used to dissect the specific functions for NSD3 in LUSC tumor initiation, progression, maintenance, and metastatic transition using conditional NSD3 gain-of-function and knockout mice. A multistep approach will also enable genetic validation of NSD3 as potential therapeutic target in advanced LUSC, a stage for which new therapies are urgently needed. In Aim 2 we will elucidate the epigenetic pathways reguated by the NSD3-H3K36me2 axis, utilizing new cutting-edge epigenomic technologies. We will also explore the role of NSD3 in promoting intratumoral heterogeneity in human and mouse models of LUSC at the single cell level. Together, this work will be the first to evaluate the therapeutic potential and mechanism-of-action of NSD3 in LUSC.

NIH Research Projects · FY 2026 · 2023-04

Identifying and targeting collateral lethal vulnerabilities in cancers Abstract/Summary Genomic deletions targeting major tumor suppressor genes frequently include adjacent passenger genes, encoding cell essential housekeeping functions. These cancer cells survive due to co-expressing functionally redundant paralogs residing in non-deleted regions of the genome. As such, these “collateral deletions” in tumor suppressor loci can confer cancer cell-specific vulnerabilities through targeted extinction of the remaining paralog. Our “collateral lethality” concept was first demonstrated in GBM with deletion of the 1p36 tumor suppressor locus encompassing ENO1, resulting in profound sensitivity to ENO2 depletion or pharmacologic inhibition (Muller et al. 2012). Subsequently, we demonstrated that deletion of mitochondrial malic enzyme 2 (ME2) in the SMAD4 locus engendered lethality upon shRNA-mediated depletion of the remaining mitochondrial malic enzyme activity encoded by the ME3 paralog (Dey et al. 2017). To systematically and comprehensively identify collateral lethal targets in cancer, we first analyzed the Broad Institute’s Cancer Dependency Map (DepMap), a publicly accessible database hosting essentiality scores of 17386 genes from a pooled genome- scale CRISPR knockout study conducted in 1054 cancer cell lines of diverse lineages (Dempster et al. 2019; Ghandi et al. 2019). The computational efforts yielded multiple collateral lethal candidate pairs including REEP3/4, PTDSS1/2, INTS6/INTS6L, PRPS1/2, LDHA/B, and CSTF2/CSTF2T via a two-class comparison method that regressed cell line sensitivity vectors against whole-genome CCLE expression and copy number data to predict paralog-depletion based sensitivity. While computational analysis of DepMap data can identify some candidate collateral lethal pairs, small sample sizes for many collateral deletions have stymied robust conclusions. Building on these computational methods, here we will apply the CRISPR/Cas12a polygenic knockout platform to systematically identify collateral lethal pairs anchored in deletion events targeting common tumor suppressor gene loci such as TP53, CDKN2A/B, ARID1A, PTEN, SMAD4, RB1 and NF1 across cancer types. This conceptual and experimental framework, coupled with a cell/organoid/tumor validation platform, seeks to identify and stringently validate collateral lethal target sets that can then be channeled into a drug discovery pipeline with the goal of expanding precision cancer treatments.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY/ABSTRACT Although mind-body interventions effectively improve psychological and physical symptoms among cancer patients, few studies have included Black cancer patients. In particular, none exists that targets Black cancer patients dealing with advanced cancer, who are vulnerable to unmanaged symptoms and report greater symptom severity and burden than their non-Hispanic White (NHW) counterparts. Prior to evaluating definitive efficacy in this population fundamental aspects regarding the feasibility and acceptability of a mind-body intervention, particularly in the area of intervention format, must be examined. For instance, while mind-body interventions are typically delivered in group-based settings, the palliative care literature has increasingly identified the need to include family caregivers in supportive care interventions as they play a central role in patients’ healthcare. This family-based approach may allow for patient-caregiver discussions pertaining to joint coping with the cancer diagnosis and also addressing caregivers’ psychological distress, which often reach clinical levels. However, such a family-based intervention may inadvertently decrease access to care, if many Black patients do not have family members who are consistent care providers or able/willing to participate jointly in the intervention. It is currently unknown whether a family-based mind-body practice is feasible and acceptable in Black patients with advanced cancer. To address these critical knowledge gaps, the objective of the proposed research is to determine the feasibility of implementing a culturally adapted mind-body intervention (Meditation- Based Support-Adapted; MBS-A) as a supportive care strategy in Black patients diagnosed with advanced cancer. The original MBS intervention includes four 60-minute sessions delivered over 4 weeks that integrates guided meditations (e.g., mindfulness, compassion, gratitude) with emotional processing techniques, which have previously been tested in predominantly NHW patient-caregiver dyads as well as in a patient group setting. To determine the intervention format for the MBS-A program (family- vs group-based) and solicit input on the original MBS intervention regarding content that needs adaptation, we will first conduct formative research that includes quantitative surveys and in-depth interviews of patients and their primary caregivers. Once the intervention format is determined and content adapted, we will conduct a randomized controlled trial (RCT) to examine the feasibility of the MBS-A intervention vs. a dose-matched attention control arm receiving a psychoeducation intervention. Participants will be assessed at baseline (prior to randomization) and 6 and 12 weeks later. Primary outcomes include indicators of feasibility regarding the overall RCT design and intervention-specific procedures. Results of this study will inform future research in which effects and mechanisms of the intervention will be tested using fully powered samples of Black advanced cancer patients to ultimately improve health in this underserved patient population.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY The rise in human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) has resulted in a rapidly increasing number of younger and healthier patients being treated with locoregional radiation therapy (RT). Although effective in curing the cancer, deleterious RT effects upon organs-at-risk (OARs) such as the salivary glands, swallowing/masticatory muscles, and mandibular bone can result in lifelong sequelae of RT normal tissue injury such as xerostomia, dysphagia, and osteoradionecrosis (ORN), respectively. These long-term sequelae in a patient population with good clinical outcomes and extended life expectancy are increasingly relevant as latent sources of morbidity and mortality in cancer survivorship. Despite this urgency, a standardized staging and monitoring system to identify patients at risk for developing ORN and other morbidities, and to assess the effectiveness of interventional therapies, is not currently available. At our institution, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is being integrated into a multimodality clinical algorithm aimed at improving diagnosis, staging, and surveillance of disease and toxicity. DCE-MRI can detect altered bone vascularity associated with bone healing, necrosis, and metastatic involvement, with excellent spatial resolution. Based on our previous NIH-funded research, we hypothesize that an increase in DCE-MRI parameters (i.e., Ktrans/Ve) at any timepoint more than 3 months after RT demonstrates increased risk of normal tissue treatment-related toxicity rates. Our objective is to conduct a biomarker analytic and clinical validation study designed to confirm DCE-MRI parameters as quantitative imaging biomarkers, with the formal deliverable of an application for FDA Biomarker Qualification at study completion, via the following specific aims: 1) Prospective qualification and validation of DCE-MRI Ktrans/Ve as an “FDA BEST-defined” quantitative imaging monitoring biomarker of ORN risk; 2) Investigation of DCE-MRI candidate biomarkers for pre-qualification as quantitative imaging biomarkers of soft-tissue structures; 3) Standardization of image acquisition and open source computational/analytic software for orodental DCE-MRI as a potential component of FDA biomarker qualification. Successful completion of this proposal has the potential to revolutionize the diagnosis and management of late RT-induced morbidities and offer clinicians a reliable and FDA Biomarker Qualification Program validated biomarker to stratify patients at risk for OARs injury after RT, monitor bone and soft tissue injury and subsequent oral morbidity, and manage post-RT care appropriately. This work is particularly relevant to NIDCR’s mission of “advancing fundamental knowledge about dental, oral, and craniofacial health and disease and translate these findings into prevention, early detection, and treatment strategies that improve overall health for all individuals and communities across the lifespan”.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Hepatocellular carcinoma (HCC) is the fastest growing cause of cancer-related deaths in the US. Immunotherapy is a promising new treatment approach for HCC, but there are numerous barriers to immunotherapy in HCC. Local intratumoral injection of immunotherapies is a logical solution to overcoming these barriers, particularly given the fact that local immune activation can drive systemic tumor immunity. However, there are substantial gaps in knowledge regarding the intratumoral delivery of immunotherapies. When delivered through conventional needles, injected medications track along the needle path and leak out into the surrounding normal tissue. Not only does this minimize treatment efficacy due to diminished on-target delivery, but it also increases systemic toxicities. Moreover, even with local deposition of immunotherapies, persistent microenvironmental barriers can inhibit the generation of tumor immunity. This application will evaluate a novel intratumoral drug delivery system with an adjustable, electrically insulating sleeve specifically designed for intratumoral delivery of immunotherapies (ImFusion system). In addition to the controlled delivery of injected drugs, ImFusion also allows for radiofrequency-mediated intratumoral hyperthermia generation. Preliminary data show that the ImFusion system results in a substantial improvement in intratumoral drug delivery relative to conventional needles, and that its hyperthermia capabilities can “prime” the tumor microenvironment for immune activation. Our central hypothesis is not only that our ImFusion needle design will improve i.t. drug delivery, but also that the hyperthermia-mediated alterations to tumor vascularity will countervail immunologic barriers and augment i.t. immunotherapy efficacy. Accordingly, the overall objectives of this proposal are to understand how 1) variations in injection technique and thermal dose influence i.t. drug deposition, 2) hyperthermia affects tumor vascularity and immune microenvironments, and 3) hyperthermia complements i.t. immunotherapy in a syngeneic rat model of HCC. We will test our hypothesis in the following specific aims: 1) Define the influence of injection technique and hyperthermia on i.t. delivery; 2) Determine the tumor microenvironmental ramifications of hyperthermia as a function of thermal dose and time; and 3) Characterize the local and abscopal effects of hyperthermia when combined with i.t. immunotherapy. The proposal is innovative because it pursues a multimodality image-guided approach to maximize tumor immunity. In doing so, the paradigm of hyperthermia as a locoregional therapy is replaced with the paradigm of hyperthermia as an “immune primer.” The proposed research is significant because it is expected to have a broad translational impact on the efficacy of immunotherapy for patients with advanced HCC.

NIH Research Projects · FY 2026 · 2023-03

Project Summary Studying regulated cell death is critical for our understanding of tumor suppression and development of novel effective cancer therapy. Ferroptosis, an iron-dependent form of regulated cell death that is induced by excessive lipid peroxidation, is morphologically and mechanistically distinct from other forms of regulated cell death such as apoptosis. However, in contrast to our deep understanding of apoptosis, how ferroptosis is regulated and coordinates with other cellular signaling in tumor suppression remains much less well understood. There also exists a significant need to translate our understanding of ferroptosis mechanisms into effective cancer therapies. Our long-term goal is to understand the mechanism(s) of action of anti-neoplastic agents and/or combinations of agents that target ferroptosis in cancer therapy. The objective of this application is to determine the role and mechanisms of AMP-activated protein kinase (AMPK), a critical sensor of cellular energy status, in regulating ferroptosis and the relevance of these regulatory functions to tumor suppression and treatment. Energy stress depletes ATP and induces cell death. Surprisingly, our recent study revealed that energy stress can potently suppress ferroptotic cell death through activating AMPK. Cancer cells with high basal AMPK activation are resistant to ferroptosis, and AMPK inactivation sensitizes such cancer cells to ferroptosis. Our recent publication and new preliminary data support the central hypothesis that AMPK inhibits ferroptosis through AMPK-mediated phosphorylation of acetyl-CoA carboxylase (ACC) and biosynthesis of polyunsaturated fatty acid (PUFA) as well as other unidentified downstream effectors. AMPK can have either tumor-suppressive or -promoting functions, depending on the context. We further hypothesize that AMPK’s tumor-promoting function is at least partly mediated by its inhibition of ferroptosis, and combining AMPK inhibitors and ferroptosis inducers (FINs) is a novel therapeutic strategy for treating AMPK-dependent cancers. To test our hypotheses, we will pursue the following specific aims: Specific Aim 1: To study the mechanisms by which AMPK inhibits ferroptosis in cancer cells. Specific Aim 2: To determine the relevance of ferroptosis to AMPK-mediated tumor development and treatment. It is expected that our proposed studies will clarify how AMPK regulates PUFA biosynthesis, identify novel regulatory mechanisms of ferroptosis pathways, and reveal a previously unrecognized function of ferroptosis suppression in AMPK-mediated tumor promotion in cancer. Our proposal is highly innovative because it focuses on a previously unexplored pathway linking AMPK regulation of ferroptosis to tumor development. Our proposed studies will have significant impact on both our basic understanding of ferroptosis and our ability to target AMPK or ferroptosis in cancer treatment.

NIH Research Projects · FY 2026 · 2023-03

Project Summary Historically, the majority of relevant research has only interrogated classical pathways in bladder cancer cells and has had little success in developing clinical drugs against bladder cancer (BC). Immunotherapy, including PD-1/PD-L1 blockade, has recently been proven effective in treating a number of tumor lineages, but the majority of BC cases are regarded as resistant or immune-quiescent tumors and are unresponsive to single checkpoint treatments. These challenges demand definition of the molecular mechanisms underlying the immuno- suppression that develops during BC progression. We demonstrated that tumor-resident Schwann cells (referred as TASc) play important roles in promoting an immunosuppressive microenvironment. TAScs express one lncRNA that modulates RAF1-mediated phosphorylation of TDO2 (Tryptophan 2,3-Dioxygenase), thereby facilitating the enzymatic activities of TDO2 and catalysis of Tryptophan to Kynurenine. The released Kynurenine in tumor microenvironment further facilitates the expansion of MDSC (myeloid-derived suppressor cells) and quiescence of effector T cells. Therefore, considering TAScs and lncRNAs as therapeutic targets may potentially sensitize BC to immunotherapy. The long-term goal of the proposal is to demonstrate the molecular mechanisms and functional importance of lncRNAs in BC so that improved strategies can be developed to reduce BC immune resistance. Our central hypothesis is that PVT1 facilitates phosphorylation of TDO2 in TAScs to promote BC immunoresistance, which could be attenuated in vivo using a targeted therapy. We will address our hypothesis from following aspects. 1) We will demonstrate the prognostic value of TAScs in BC and determine the functional importance of TASc expressing lncRNA in BC tumorigenesis (Aim 1). We will determine the underlying molecular mechanisms of lncRNA in regulating the enzymatic activities of TDO2 and the IL-6 induced, RAF1-mediated phosphorylation of TDO2 (Aim 2). 3) We will ascertain the functional importance of TAScs using small molecule inhibitor and small molecule inhibitor-conjugated anti-sense oligonucleotides, anti-IL-6 neutralization antibody, or kynurenine aminotransferase inhibitor in combination with immunotherapy in inhibiting BC immune resistance and metastasis (Aim 3). Emerging evidence of the oncogenic involvement of lncRNAs, as well as their implicated roles in mediating immunosuppression, warrants further characterization of TASc-specific lncRNAs and future applications that hinge on their activity. Our goal is to demonstrate the underlying mechanisms of BC immune resistance from lncRNA and metabolite points of view. Thus, a strategy that combines immune checkpoint inhibitors and lncRNA- based therapeutic strategies has the potential to significantly advance BC treatment. In the long run, these research findings will benefit the cancer community by introducing the robust clinical effects of targeting TAScs and TASc-expressing lncRNAs as promising therapeutic targets.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY / ABSTRACT Regular physical activity plays an important role in disease prevention and helps people live longer, feel better, and perform daily tasks more easily. Yet, less than 25% of American adults meet physical activity recommendations, and Americans spend an average of 9.5 hours per day sedentary, putting them at increased risk of cardiovascular disease, cancer, and mortality. Black and rural adults are less likely to be physically active and more likely to be sedentary than non-Hispanic White adults and those in urban areas, contributing to racial and ethnic and rural health disparities. Interventions focused on increasing moderate-to-vigorous physical activity have had limited success among Black adults residing in urban and rural areas. Reducing sedentary behavior as a means to increase physical activity may be a more effective approach. Yoga is a practice that combines the mind, body, and spirit through light-intensity movement, breath awareness, and relaxation techniques, and can be a valuable tool to reduce sedentary behavior and promote movement in adults who are insufficiently active and need to build up to regular physical activity. Furthermore, as a mindfulness practice, yoga’s effects on stress may be particularly relevant to Black adults who experience unique stressors, including racism and discrimination. Based on this, we partnered with churches to develop and test the feasibility of Harmony & Health (HH), a theoretically based intervention that integrates yoga practices with Christian spirituality to promote psychosocial wellbeing and movement among insufficiently active Black adults. We first tested the feasibility of HH among Black adults residing in an urban city, and then tested the feasibility of HH among a diverse sample of rural adults. This study is the next logical step and expands on our pilot studies that confirmed the feasibility of HH in Black adults. Feasibility and acceptability were high as measured by study completion (≥80% across studies), attendance (M urban=10.5±3.7; M rural=10.6±4.2), and participant satisfaction (≥97.2% overall across sites). HH also improved psychosocial wellbeing and reduced sitting time. The proposed study builds on these efforts by randomizing 100 Black adults who are not meeting physical activity recommendations from two geographically distinct sites, one urban and one more rural, to participate in HH or an attention control condition. HH sessions will take place twice a week for 8 weeks and focus on sitting less and moving more using stretching, breathing, and guided relaxation with Christian scripture. Assessments occur at baseline (week 0), post-intervention (week 9), and follow up (week 24). Results of this study will determine our ability to deliver these interventions with strong rigor and fidelity across multiple sites and will establish the feasibility and acceptability of this intervention among urban and rural Black adults. Findings from this study will be used to modify training and implementation procedures in preparation for a future fully powered multi-site randomized controlled efficacy trial. If supported, HH could be an innovative strategy to improve psychosocial wellbeing and physical activity in low active Black adults.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Dysphagia (difficulty swallowing) is a highly prevalent and impactful condition with significant burden on the healthcare system. Across the lifespan, dysphagia is associated with excess risk of mortality, increased length of stay, aspiration pneumonia, and malnutrition thereby elevating medical costs and resource utilization. Not only a health problem, dysphagia also adversely affects quality of life and daily function with disproportionate impact on cancer survivors. Adoption of evidence-based methods into clinical practice lags decades behind discovery. One such gap is adoption of evidence-based practices (EBP) by speech-language pathologists in dysphagia management. Evidence-based dysphagia care begins with evidence-based swallowing evaluation. Significant progress has been made in the field of dysphagia to develop evidence-based evaluation methods, with particular emphasis on physiologic characterization of swallowing. The relative safety and efficiency of swallowing, that is how well a food or liquid bolus is kept out of the airway and clears fully through the pharynx into the esophagus, is a fundamental driver of clinical decision making – yet, remains inconsistently assessed and reported in clinical practice. To address this gap, the investigators’ developed DIGEST™ (Dynamic Imaging Grade of Swallowing Toxicity). DIGEST is an EBP tool to grade the severity of pharyngeal dysphagia based on results of a radiographic (videofluoroscopic) modified barium swallow (MBS) study. DIGEST uses a basic flowsheet and rubric (available open access via PMC) to summarize the patterns of penetration/aspiration and pharyngeal residue observed on the MBS as markers of swallowing safety and efficiency. DIGEST is a pragmatic yet robust measure validated in the head and neck cancer population, and adopted into routine practice at the PI’s institution with over 11,000 MBS graded in the clinic using the methodology since development in 2016. Peer-reviewed research shows adoption of DIGEST in external academic medical settings and federally funded clinical trials. Despite this promise, several obstacles still limit widespread adoption in routine cancer care. These include scalability to fit diverse clinical contexts outside the PI’s environment and uncertainty about best implementation strategies. The long-term goal of this project is to improve dysphagia care and patient outcomes through reliable adoption of DIGEST into routine clinical practice. Our central hypothesis is that DIGEST scales-up maintaining validity in diverse cancer populations under common clinical practice variations with reliable adoption facilitated by an active implementation strategy. The objective of this application is to use dissemination and implementation (D&I) science to accomplish the following Specific Aims: 1) demonstrate validity of DIGEST in diverse oncology populations and imaging acquisition protocols, 2) examine context and fidelity of natural dissemination of DIGEST in real-world, early adopters, and 3) evaluate active implementation strategies to improve reach and fidelity of DIGEST in clinical practice. With dense multi-site networks and content expertise, the investigators are uniquely equipped to conduct the proposed D&I project. We expect this work to improve care by narrowing the research-to-practice gap in dysphagia diagnostics.

NIH Research Projects · FY 2026 · 2023-02

Summary In 2020, breast cancer has surpassed lung cancer as the most commonly diagnosed cancer in women. Compared to estrogen receptor (ER)-positive breast cancers, ER-negative (ER-) breast cancers have worse prognoses and no effective prevention strategies. In this study, we will explore new strategies for immunoprevention of ER- breast cancer. Inducing potent anti-tumor immunity for prevention of poorly immunogenic breast cancers has been highly challenging. Engagement and expansion of activated dendritic cells (DCs) could facilitate broad and efficient anti-tumor immunities. However, certain existing DC stimulators (e.g., agonists of toll-like receptors and STING) also triggered adverse immune responses. For cancer prevention, it is imperative to develop safe and effective approaches to boost DC immunity. To this end, we screened for dietary supplements that increase DC activities and identified natural vitamin E (VitE) as a stimulator of DC functions. Excitingly, we found that breast cancer patients who took VitE during immunotherapies had a significantly better survival rate and improved therapeutic response than patients who didn’t take VitE, suggesting that VitE administration may potentiate anti-tumor immunity. Indeed, systemic (oral) administration and local (at injection site together with cancer vaccines) delivery of VitE significantly prolonged tumor-free survival in ER- mammary tumor mouse models that didn’t respond to cancer vaccines alone. These data led us to hypothesize that VitE administration, via reinforcing DC activation and antigen presentation, enhances immunoprevention of ER- breast cancer by cancer vaccines. We will test whether VitE could enhance cancer vaccine-induced immune surveillance and prevent/delay tumor initiation/progression in genetic engineered mouse models of (HER2+ and basal-like subtypes) ER- mammary tumors and the CD34+ humanized mouse models (for prevention of human ER- breast cancers) (Aim1). As a proof of concept, we will primarily use a triple- antigen (tumor associated antigens neu/IGFBP-2/IGF-IR) peptide vaccine (TAVac) for proposed studies since TAVac has shown partial efficacy in delaying tumor progression in ER- mammary tumor mouse models. Importantly, corresponding DNA vaccines against human HER2/IGFBP-2/IGF-IR are currently under phase I/II clinical studies for prevention of HER2+ and HER2- breast cancer recurrence. To gain mechanistic insights into how VitE potentiates anti-tumor immunity, we will investigate i) the global effect of VitE on the immune cell landscape by mass cytometry (CyTOF); ii) the impact of VitE on DC and T-cell subset compositions, functionality and signaling networks; iii) major immunophenotype changes critical for VitE-enhanced immunoprevention; iv) how VitE prompts antigen processing/presentation in DCs and whether VitE functions through SHP1, a critical DC checkpoint (Aim2). Finally, we will test novel strategies to further improve the immunoprevention efficacy against ER- mammary tumors (Aim3). If successful, our strategies could be readily tested in future clinic trials for immunoprevention of breast cancer, particularly, for women at high risk for ER- breast cancer.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT Immune checkpoint therapy (ICT) has revolutionized the care of several malignancies, resulting in durable response and even cure in a small subset of cancer patients. However, unlike tumors that are highly responsive to ICT (e.g., melanoma), the majority of patients with advanced prostate cancer respond poorly to ICT and no survival benefits have been observed in non-selective patients. Although recent preclinical and clinical studies have shed some light on the mechanisms of immunoresistance in prostate cancer, the cellular and molecular basis of immunoresistance in prostate cancer remains poorly characterized. Myeloid-derived suppressor cells (MDSCs), a group of pathologically activated monocytes and neutrophils with potent immunosuppressive activities, have been implicated as one of the key mechanisms in driving tumor progression, metastasis, and therapeutic resistance, including resistance to ICT, in various cancers. MDSCs can be classified as polymorphonuclear or granulocytic MDSCs (PMN-MDSCs or G-MDSCs) or monocytic MDSCs (M-MDSCs), with PMN-MDSCs as the predominant population in most cancer types. We and others have demonstrated that PMN-MDSCs are the major MDSC subset in mouse and human prostate cancers, playing an important role in prostate cancer progression and resistance to anti-androgen therapy and ICT. Correspondingly, therapeutically targeting MDSCs delays prostate cancer progression and improves responses to anti-androgen therapy and ICT in preclinical models. Importantly, PMN-MDSCs may be clinically relevant to prostate cancer, as they are abundantly present in both primary and metastatic tumors. Emerging evidence suggests that metabolic reprogramming of PMN-MDSCs plays an important role in their immunosuppressive activities, yet the underlying molecular mechanisms are still poorly defined. Through transcriptome analyses (single-cell RNA-seq, microarray, and bulk RNA-seq) of multiple datasets, we identified Acod1, which encodes cis-aconitate decarboxylase (ACOD1), as one of the most highly expressed metabolic genes in immunosuppressive PMN-MDSCs. ACOD1, which catalyzes the synthesis of itaconate from cis- aconitate in the tricarboxylic acid (TCA) cycle, is a novel immunomodulator with potent anti-inflammatory and antimicrobial effects in mammalian cells, especially in macrophages. Our results unexpectedly showed that ACOD1 may also be a potential regulator of PMN-MDSCs. We hypothesize that ACOD1 promotes tumor progression and resistance to ICT in prostate cancer through metabolic reprogramming of PMN-MDSCs. We will test our hypothesis in the following aims: Aim 1). Determine the role of ACOD1 in regulating the immunosuppressive activities of PMN-MDSCs. Aim 2). Determine the role of ACOD1 in PMN-MDSCs in driving prostate cancer progression. Aim 3). Determine whether Acod1 KO improves the response of prostate cancer to ICT.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT More than 60% of metastatic castration-resistant prostate cancers (CRPC) contain PTEN and TP53 gene deletions and mutations. To date, there are limited treatment options for this molecular subtype. The overall objective of this application is to elucidate the complex role of B7-H3 signaling in prostate cancer (PCa) as well as develop biomarker-driven B7-H3 targeted therapy for CRPC. Our preliminary studies demonstrated that PTEN/TP53 defects induce B7-H3 overexpression and that depletion of B7-H3 in cancer cells remarkably impaired progression of PTEN/p53-deficient PCa in vivo. Mass cytometry analysis revealed that B7-H3 signaling is involved in modulating immunosuppressive myeloid cells in the PCa tumor microenvironment (TME). Here, the central hypothesis is that B7-H3 signaling contributes to the progression of PCa containing PTEN/p53 deficiencies and plays a key role in reprogramming immunosuppressive myeloid cells in the PCa TME. The central hypothesis will be tested by pursuing the following specific aims. Aim 1. Elucidate B7-H3’s impact on progression of advanced PCa containing PTEN/TP53 loss. The hypothesis in this aim is that B7-H3 plays a key role in PCa progression and resistance to ADT and that PTEN/TP53 defects render CRPC more responsive to B7-H3 targeted therapy. To this end, we have generated a novel GEMM with prostate-specific co-deletion of Pten/Trp53/Cd276. This state-of-the-art spontaneous GEMM provides a unique tool to dissect the role of B7-H3 signaling in PCa development and therapeutic resistance. Mechanistic studies will illuminate how PTEN and p53 pathways control B7-H3 expression. Clinical relevance among B7-H3 expression, PTEN/TP53 status, and disease progression will be assessed in human CRPCs. B7-H3 targeted therapies will also be tested in various preclinical models of metastatic CRPC containing PTEN/TP53 loss. Aim 2. Understand how B7-H3 signaling modulates immunosuppressive myeloid cells in CRPC. The hypothesis is that B7-H3 signaling contributes to the reprogramming of myeloid cells in the PCa TME. Single-cell transcriptomics analysis will be performed in above GEMMs. Functional studies will address how B7-H3 signaling mediates the crosstalk between cancer cells and immunosuppressive myeloid cells. B7-H3 receptor(s) and the downstream signaling pathway(s) in myeloid cells will also be characterized. Finally, the therapeutic potential of co-targeting B7-H3 and immunosuppressive myeloid cells will be evaluated in preclinical models of CRPC. These studies are expected to have significant positive impacts, including advancing the understanding of B7-H3 biology in cancers and providing a compelling rationale for the use of PTEN and TP53 defects as molecular biomarkers for predicting response to B7-H3-targeted therapy for patients with CRPC. The proposed research will also establish, for the first time, the role of B7-H3 signaling in reprogramming myeloid cells in the TME and develop effective combinatorial immunotherapy targeting B7-H3.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT Iron is vital for many physiological processes, but excessive iron causes toxicity. Dysregulated iron homeostasis (either iron deficiency or overload) is a harbinger of pathological conditions. The liver stores iron in hepatocytes and is the major organ that controls systemic iron homeostasis. Liver cancer, primarily hepatocellular carcinoma (HCC), is highly lethal with limited treatment options and no biomarkers to predict therapy response. Leukemia inhibitory factor receptor (LIFR) is frequently downregulated in human HCC; however, in vivo and genetic studies of LIFR’s functions in liver cancer development and therapy response were lacking. Recently, by constructing and characterizing hepatocyte-specific and inducible Lifr-knockout mice, we found that loss of Lifr promoted liver tumorigenesis and conferred resistance to sorafenib-induced ferroptosis, a non-apoptotic type of cell death characterized by the iron-dependent accumulation of lipid hydroperoxides. Our data also pointed to a role for LIFR in inhibiting NF-κB signaling in the liver, which in turn downregulates lipocalin 2 (LCN2), an iron- sequestering cytokine. In parallel, our data revealed that in oncogene-induced liver tumors, overexpression of LIFR increased, while knockout of Lifr decreased CD8+ T cell infiltration, which may be mediated by LCN2- dependent downregulation of iron levels, viability, and proliferation of T cells. Altogether, these data support a hypothesis that loss or downregulation of LIFR in liver cancer leads to upregulation of LCN2, which on one hand confers resistance to ferroptosis on liver tumor cells, and on the other hand, deprives T cells of iron that is essential for T cell viability, proliferation, and effector function; both mechanisms contribute to liver cancer progression and therapy resistance. In the proposed work, we will elucidate the molecular mechanisms by which LIFR inhibits NF-κB signaling in liver cells (Specific Aim 1). Further, we will investigate whether LCN2 can serve as a therapeutic target for enhancing sorafenib efficacy in HCC (Specific Aim 2). Finally, we will study whether LIFR or therapeutic LCN2 neutralization can sensitize HCC to immunotherapy (Specific Aim 3). Genetically engineered mouse models, Sleeping Beauty transposon-mediated oncogene-induced liver cancer models, and HCC patient-derived xenograft models will be used to study the therapeutic potential and mechanisms of action of two novel drug combinations, which will illuminate how to improve liver cancer therapy by targeting an iron- sequestering pathway. We envision that low LIFR expression and high LCN2 expression could be used to select HCC patients who will likely benefit from the combination therapy with the LCN2-neutralizing antibody plus sorafenib or immune checkpoint inhibitors.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY DNA topoisomerases are types of enzymes that can specifically resolve topological stresses by transiently introducing strand breaks into DNA molecules and enabling the rotation of the supercoiled DNA strand. Mammalian cells encode two types of topoisomerases: type I topoisomerases (TOP1, TOP1mt, TOP3A, and TOP3B), which introduce single strand breaks into DNA, and type II topoisomerases (TOP2A, TOP2B, and SPO11), which introduce double strand breaks (DSBs) into DNA. This proposal focuses on type II topoisomerases, i.e. TOP2A/2B, in human somatic cells. During cleavage reaction, the tyrosine in the catalytic active site of TOP2 is covalently linked to the DNA backbone and forms the so-called topoisomerase II cleavage complex (TOP2cc). Under normal conditions, TOP2cc forms transiently and is not detectable. However, a wide variety of topoisomerase poisons, including etoposide, have been developed and used as chemotherapeutic drugs for cancer treatment. Mechanistically, etoposide acts to stabilize TOP2cc, which eventually lead to DNA strand breaks and kill tumor cells. While many investigators including us investigated TOP2-induced DNA lesions and how they can be repaired by different repair pathways, this proposal focuses on a new concept that cells have evolved distinct pathways to avoid and limit DNA lesions induced by TOP2. In this proposal, we will determine mechanistically how several unique TOP2 regulators act together to avoid DNA damage and therefore promote cell survival. Results from these studies are critically important for the understanding of therapeutic response to etoposide and other anti-cancer agents. .

NIH Research Projects · FY 2026 · 2023-01

Cancer immunotherapy (IMT) can produce robust and durable anti-tumor immune responses in some cases. However, many cancers are non-responsive to IMT and combination approaches need to be actively investigated, particularly in lethal tumors such as IMT-insensitive non-small cell lung cancer (NSCLC). Preclinical studies in general have been found to be poor predictors of success for IMT agents and chemoradiotherapy combinations in the clinic, likely due to poorly conceived and executed treatment protocols, dated disease model systems and lack of an existing framework for cross-validation of preclinical results. There is a need to develop a rigorous preclinical testing program for existing IMT agents combined with chemoradiation. NSCLC genetically engineered mouse models (GEMMs) of the major molecular NSCLC subtypes have been created. However, there are no NSCLC GEMMs that to our knowledge has demonstrated “abscopal” responses reliably to IMT, which is one unique strength of this current proposal. Also, a major limitation of existing GEMMs is the relatively small number of different genotypes that can be generated and their lack of quantitative precision. This proposal leverages a new technique, tumor barcoding with barcode deep-sequencing (Tuba-seq) and in vivo Cre-lox and CRISPR/Cre-mediated GEMMs to model oncogenesis and radiation-drug response with unprecedented precision and genomic-comprehensiveness. We are using this R01 mechanism in the present proposal via two Specific Aims stated below to test the following central hypotheses: (i) we hypothesize that treating with both anti-PD-L1 and a novel orally bioavailable ATR inhibitor (ATRi), AZD6738, in combination with chemoradiation will result in an improved and durable anti-tumor immune response in poorly immunogenic NSCLC GEMMs; and, (ii) Tuba-seq we will allow an unprecedented view of the radio-pharmacogenetic landscape of NSCLC responses in vivo. SPECIFIC AIM #1 – Establish a radio-pharmacogenetic map of oncogene-driven non-small cell lung cancer to both chemoradiation and combined chemoradiation/anti-PD-L1 therapy We propose to use novel CRISPR/Cre-mediated GEMMs of NSCLC to test the optimal combinations of chemoradiation with anti-PD-L1 IMT. We will then examine the mechanism of action of chemoradiation with IMT using multiparametric immunologic approaches and an innovative technique enabling lineage tracing and direct quantification of treatment effects on these different genetic backgrounds in vivo. SPECIFIC AIM #2 – Determine tumor cell genotype effects on combination anti-PD-L1 immunotherapy and ATRi with chemoradiotherapy in oncogene- driven non-small cell lung cancer. This Aim leverages our novel CRISPR/Cre-mediated GEMMs of NSCLC to test genotype effects on the combination of anti-PD-L1 and ATRi with chemoradiation. By using all these tools, we will be able to decode the major aspects of the molecular underpinnings of chemoradiation and combination IMT resistance in NSCLC, contributing to improved therapies and positively impacting patient outcomes.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDAC) contains a desmoplastic stroma that limits blood perfusion and thus the delivery of nutrients, oxygen, and therapeutics, creating a hypoxic microenvironment that is able to resist nearly all forms of treatment, including immunomodulating therapy. Cancer-associated fibroblasts (CAFs) are the main components and producers of stroma in PDAC. The hypoxia-inducible factors-1 (HIF1) and -2 (HIF2) are stabilized in low oxygen and have been hypothesized to mediate therapeutic resistance and aggressive growth of PDAC, but deletion of HIF1 or HIF2 in the pancreatic epithelial compartment produced no obvious phenotype in PDAC. Thus, we reasoned that HIF may instead be supporting PDAC pathobiology through its functions in cancer-associated fibroblasts (CAFs), which are the other major cellular component in pancreatic tumors. Since little was known about the role of HIF in the stromal compartment, we created a mouse model that exploits two recombinases enabling us complete spatiotemporal control of tumor growth and HIF ablation in CAFs. Using this model, we found that the abrogation of HIF2 in CAFs decreased tumor growth and doubled the median survival of animals with pancreatic cancer. The loss of HIF2 in CAFs correlated with fewer intratumoral M2 macrophages and regulatory T cells. Furthermore, conditioned media from hypoxic CAFs produced similar effects in ex vivo assays even after boiling. These preliminary data suggested a HIF2- dependent crosstalk between CAF and the immune compartment that may be critical to the ability of PDAC to evade the immune system. Building on this preliminary data, we propose to identify the molecular mechanisms that allow hypoxic CAFs to modulate the immune microenvironment of pancreatic cancer through three aims. In Aim 1, we will determine the molecular mediators of HIF2-dependent crosstalk between CAFs and macrophages. We will use mass spectrometry to identify the HIF2 dependent factor secreted from CAFs capable of inducing M2 polarization in macrophages and determine how this factor is produce in CAFs and how it alters macrophage function. Aim 2 will define the HIF2-dependent relationships between CAFs and T lymphocytes. We will explore how CD8+ suppression and Treg conversion is regulated by HIF2 signaling in CAFs using mouse models and co-culture methods. Lastly, in Aim 3, we will repurpose HIF2 inhibitors from their current indications in renal cell carcinoma to pancreatic cancer as a drug to enhance immunotherapy responses. The proposed research is significant because pancreatic cancer does not respond to immunotherapy, and our data suggests that HIF2 inhibition may greatly improve immune responses. This research is innovative because we will provide the first description of the immune-targeting molecular pathways activated by hypoxic microenvironment in PDAC. Further, we will offer the first mechanistic evaluation of a potential therapeutic combination using a HIF2 inhibitor along with immune checkpoint blockade as the first steps towards clinical translation.

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract Premature senescence-triggered vascular diseases (PmSVD) induced by ionizing radiation (IR), as well as Hutchinson-Gilford progeria syndrome (HGPS), are notably characterized by accelerating processes of atherosclerosis (AthS) and coronary artery disease (CAD). Although endothelial dysfunction in PmSVDs is well known, there is a paucity of available treatments to prevent PmSVD-induced CAD; hence, there is an urgent need to fill this gap. Persistent senescence-associated secretory phenotype (PISP), provoked by TL dysfunction, plays a central role in cancer recurrence and resistance, but its regulatory mechanisms and contribution to AthS remain unknown. Our long-term goal is to determine the molecular mechanisms by which PmSVD induces PISP in endothelial cells (ECs) and CAD. PmSVD significantly up-regulated TOP2β degradation via PKCζ activation. The depletion of EC TOP2β instigated PARP activation and PISP; it also accelerated AthS. We showed the critical role of mtROS in PKCζ activation, which is one of the initial steps for the Mt-nucleus feedback loop. Of note, the crucial role of mtROS in both IR and HGPS has been well established. Lastly, by performing IC-MS analysis in both IR and HGPS ECs, we also found that the following 3 metabolite-related pathways were regulated in IR and HGPS ECs in common: 1) nucleotide sugars-glycosaminoglycans (GAGs) and sulfate, 2) glutamate, and 3) NAD+- hydrogen sulfide (H2S). Although the contribution of all 3 metabolites pathways to CAD and aging has been suggested, the exact role and mechanical insights in regulating PmSVD remain largely unknown. We propose the novel hypothesis that PmSVD-induced mtROS activates the PKCζ-TOP2β module, followed by TOP2β degradation, and instigates TL DNA damage. TL DNA damage promotes PARP activation, which induces mt dysfunction and forms an mt-nucleus feedback loop, resulting in persistent metabolites changes, including nucleotide sugars and NAD+-H2S pathways, causing PISP and CAD. We will test our hypothesis by pursuing the following 3 specific aims: In Aim 1, we will determine the role and regulatory mechanisms of the following 3 common metabolites-related pathways in PmSVD in vitro; 1) nucleotide sugars-GAGs and sulfate, 2) glutamate, 3) NAD+- H2S. in Aim 2, we will characterize the role of PKCζ-TOP2β module and PARP1 in PmSVD-mediated metabolites changes and mt dysfunction in vitro. In Aim 3, we will determine the role of the PKCζ-TOP2β module and subsequent PARP activation in PmSVD-induced coronary AthS (CAthS) in vivo. The proposed work is expected to establish the roles of PKCζ-TOP2β and PARP as the hub molecules in regulating PmSVD-induced metabolite changes and PISP. The approach is innovative because we will use the new technologies of iPSC, ion chromatography-mass spectrometry (IC-MS), machine learning, imaging mass cytometry, and a novel mouse CAthS model. The proposed research should positively impact PmSVD by leading to a novel approach to inhibiting PISP.

NIH Research Projects · FY 2026 · 2022-12

ABSTRACT KRAS is one of the most frequently mutated genes in human cancers. Despite advances in the development of inhibitors that directly target mutant KRAS and the FDA approval of KRASG12C inhibitor sotorasib for KRASG12C- mutant non-small cell lung cancer (NSCLC), cancer cell adaptation and resistance to KRAS inhibitors are almost inevitable and remains a major challenge that limits their clinical benefits. Our preliminary data establish proteostasis reprogramming as an essential mechanism that mediates tumor resistance to KRAS inhibitor. Inactivation of oncogenic KRAS rapidly downregulates both the heat shock response (HSR) and IRE1a branch of the unfolded protein response (UPR). However, only IRE1a is selectively reactivated in KRASi-resistant tumors. Genetic or pharmacologic suppression of IRE1a substantially sensitizes KRASG12C-mutant tumors to sotorasib, leading to complete and durable responses in preclinical NSCLC and pancreatic cancer models. Mechanistically, we found that oncogenic KRAS-MAPK signaling promotes IRE1a protein stability through direct ERK-IRE1a interaction. In contrast, multiple mechanisms of resistance to KRASi, including reactivated ERK and hyperactivated AKT, converge to re-activate IRE1a in resistant tumors. These findings provide a framework to seek biological insight into the proteostasis reprogramming in KRAS-mutant cancers, and to further explore the effects of pharmacological inhibition of proteostasis reprogramming as an anti-tumor approach for KRAS-mutant cancers. We hypothesize that IRE1a-mediated proteostasis reprogramming facilitates tumor resistance to oncogenic KRAS inhibition and that multiple resistance pathways converge with IRE1a to restore proteostasis and promote therapy resistance to KRAS inhibitors. This proposal will determine the molecular mechanisms of differential IRE1a regulation in response to mutant KRAS inhibition (Aim 1), define proteostasis machinery crosstalk between HSR and UPR in KRAS-mutant cancers (Aim 2), and evaluate the therapeutic efficacy of targeting proteostasis reprogramming to overcome KRASi resistance in KRAS-mutant cancers (Aim 3). Accomplishing these aims will establish the biological significance and biochemical basis of oncogenic signaling regulated proteostasis network in KRAS-mutant human cancers, leading to development of more effective and well-tolerated therapeutic strategy to reverse KRASi resistance and bypass the on-target toxicity of targeting multiple resistance signaling pathways.