University Of Arizona

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY: Immunosuppressed transplant recipients have a 65-253 fold higher risk of developing cutaneous squamous cell carcinoma (cSCC) and are contraindicated for treatment with immune checkpoint inhibitors, presenting an important unmet clinical need. The ability of T cells to constrain cSCC is demonstrated by the response of 32- 46% of immunocompetent patients to immune checkpoint inhibitors. However, cSCC has the potential to evade an active T cell response as demonstrated by the formation of cSCC in immunocompetent patients and resistance to immune checkpoint inhibition in some patients. Prior work suggests that evasion of an active T cell response occurs through the process of immunoediting, in which T cells destroy tumors that present mutated tumor proteins that bind the T cell receptor (neoantigens), and thus select for less immunogenic tumors. This proposal will compare the neoantigen profile and immune escape mechanisms in tumors and tumor-adjacent skin from immunosuppressed and immunocompetent individuals as a novel approach to evaluate the role of T cells in immunoediting. Evaluating the neoantigen profile in carcinogen-exposed tumor- adjacent skin will additionally provide evidence for immunoediting before the formation of a clinically apparent lesion. Furthermore, since cSCC in immunosuppressed patients develops in the context of diminished T cell function, this proposal tests the innovative concept that these patients will have a neoantigen profile that is more amenable to treatment with a personalized neoantigen vaccine. The Hastings laboratory has created an MHC class I neoantigen prioritization model with high accuracy in predicting neoantigens that elicit a T cell response, which will be applied to evaluate the neoantigen profiles of cSCC from immunosuppressed and immunocompetent individuals. The Hastings laboratory has also generated and characterized a solar- simulated light induced, transplantable cSCC tumor that will be used to test vaccine efficacy in the proposed studies. Preliminary data demonstrate that the cSCC transplantable model is constrained by T cells and vaccination with irradiated tumor cells protects from tumor challenge. This proposal will test the central hypothesis that cSCC and carcinogen-exposed, tumor-adjacent skin from immunosuppressed individuals will have a more immunogenic neoantigen profile and less frequent immune escape mechanisms compared to cSCC from immunocompetent individuals. Aim 1 of this proposal will compare the neoantigen profile and immune escape mechanisms between immunosuppressed and immunocompetent patients. Aim 2 will compare the neoantigen profile and immune escape mechanisms of cSCC from mice with and without a functional T cell repertoire and demonstrate the efficacy of cancer vaccines in immunosuppressed mice. The impact of the project is to provide evidence for neoantigen vaccines as an important treatment option for immunosuppressed patients and systematically characterize the immune escape mechanisms in cSCC to determine additional targets of therapy for cSCC in immunocompetent patients.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY / ABSTRACT This application, entitled “Student Transformative Experiences to Progress Undergraduate and Graduate Professionals” (STEP-UP) for cancer prevention, is a multidisciplinary research training initiative led by the Health Promotion Sciences faculty and the University of Arizona (UA) Comprehensive Cancer Center, Cancer Prevention and Control (CPC) Program. It is uniquely designed to provide 75 upper division undergraduate and 25 Masters' degree seeking students with an intensive, summer research experience in CPC over the grant period. The research training includes immersion in mentor research laboratories and programs full-time for 10 weeks to experience CPC science across the entire continuum of basic to applied and even dissemination research. Faculty mentor matching is carefully tailored to trainee interests, gaps in prior training, and career goals, from 39 faculty members from diverse disciplines committed to this program who are funded in CPC research. Trainees will also engage in weekly complimentary educational activities, including learning about community engagement and career exploration and development. Educational pedagogy will inform on our methodology in order to assure students receive a quality educational experience that not only will increase understanding, but also will motivate students toward a continued educational commitment to advance in CPC sciences. An embedded mentor development program will enhance faculty mentor training and engage Masters' students and high-performing STEP-UP trainees, from past years, to serve as peer-to-peer mentors. The STEP-UP program fills early gaps in our research training pathway programs, builds on purpose-driven efforts at the UA Cancer Center to attract and retain a diverse student body in CPC research, leverages our international reputation in CPC, supports continuous renewal of CPC mentors in academics and care through the mentor training program, and has garnered significant institutional support. Our specific aims are: 1. Recruit and retain undergraduate and Master's level graduate students into the STEP-UP research training program; 2. Provide a multidisciplinary research training experience in CPC that strengthens self-efficacy and intrinsic motivation to become a CPC scientist; 3. Support mentor training across academic stages, from mentee to peer-to-peer up through senior faculty mentors, and cultivate professional relationships with scientists, research programs, and community partners; 4. Conduct formative and summative evaluations to improve the program over time. The training program is centralized in Southern Arizona and offers hands-on research training within our unique catchment area which is rich in diversity relative to ethnicity (31.7% Hispanic), race (5.3% Native Americans) and age (18.0% ≥65 years). This distinctive location, strong community ties, university facilities and resources for research, as well as committed and experienced faculty mentors and leaders, assures a quality program that will build and sustain the CPC workforce of the future.

NIH Research Projects · FY 2026 · 2023-07

Irritable bowel syndrome (IBS) affects an estimated 10-15% of the U.S population and induces morphologic and physiological abnormalities significantly impairing one’s quality of life and is the most common diagnosis of a heterogeneous group of gastrointestinal disorders of gut-brain interaction (DGBI). The risk of IBS following an acute gastrointestinal (GI) infection is approximately 9%, and has been linked to numerous bacterial, protozoan, and viral infections. Notably, SARS-CoV-2 infection elicits a wide range of GI symptoms, including diarrhea, nausea, and vomiting, with reports of acute GI symptoms occurring in up to 61% of patients. Initial studies have shown persistent GI symptoms lasting up to 5-6 months post-acute infection in 40-44% of SARS-CoV-2 patients. Given the scale of the ongoing pandemic and reports of chronic GI symptoms after acute SARS-CoV-2 infection, determining how this pathogen will impact the incidence or exacerbate IBS symptoms, while playing a major role in the development of post-acute SARS-CoV-2 (PASC), known colloquially as Long COVID, is imperative. However, to date, there is a dearth of studies that have assessed the development of post-acute GI disorders following SARS-CoV-2 infection. The Arizona CoVHORT, an ongoing, prospective, longitudinal study of the acute and long-term impacts of SARS-CoV-2 infection on adults, provides the critical extant infrastructure required to efficiently investigate the health impacts of the pandemic. Using this cohort infrastructure, we propose the following aims: (1) Estimate the incidence of IBS following SARS-CoV-2 infections compared to non- infected participants. To determine the incidence of IBS following SARS-CoV-2 infection, we will employ data from the Rome IV IBS diagnostic questionnaire to compare rates of new onset IBS among participants who tested positive for SARS-CoV-2 to those who did not, while controlling for confounding factors such as age, gender, and ethnicity comorbidities and concomitant stress at the time of infection. (2) Determine the role of pre-existing IBS on the development and severity of PASC. We will follow IBS participants who reported a diagnosis (1) prior to March 2020, (2) before a SARS-CoV-2 infection, and (3) those who report no history of infection to determine their ongoing and long-term symptoms over 2-5 years, including assessment of risk factors and confounders. (3) Establish mechanisms of IBS following SARS-CoV-2 infections including differences in the fecal microbiome composition and function, the host’s anti-commensal immune response to the fecal microbiome, and targeted/untargeted serum protein biomarkers among SARS-CoV-2 exposed and unexposed, who do and do not develop incident IBS. We will collect blood and stool samples and employ shotgun metagenomics, host-microbiome directed IgG-seq and IgA-seq, and high dimensional serum proteomic arrays to explore novel mechanisms, phenotypes, and biomarkers associated with PASC-IBS. PASC will impact the individual health of millions of Americans over the next several years, and to date, limited studies have examined potential long-term effects of SARS-CoV-2 on GI outcomes specifically.

NIH Research Projects · FY 2026 · 2023-06

Despite recent advances in the understanding host-microbiome interactions in the pathogenesis of Inflammatory Bowel Diseases (IBD), the complexity of the host’s response to changing gut microbiota is daunting and still incompletely understood. In this proposal, we show that poly-ADP-ribosylation (PARylation), a post-translational modification that involves the enzymatic transfer of ADP-ribose (ADPr) from NAD+ to specific amino acids of target proteins, plays key roles as a mediator of inflammatory response in the gut. For the first time, we provide evidence that PARP1 is the primary PAR writer in the colon, where it serves as a powerful transcriptional modulator. Commensal bacteria are necessary for mucosal PARP1 activity and PARylation and, reciprocally, PARP1 controls the microbial composition and metabolic activity, modulates colonic epithelial barrier function, and restricts the mucosal Treg compartment. Importantly, human and murine colitis is associated with mucosal hyperPARylation, which can be transferred to germ-free mice with complex microbial community from IBD patients. Total or epithelial-specific knockout of PARP1 (or pharmacological inhibition) protect from and promote recovery from mucosal injury. Based on these novel preliminary observations, we hypothesized that hyperPARylation is a significant contributor to mucosal inflammation and impaired mucosal restitution via both extrinsic (interaction with gut microbiota) and intrinsic effects in the colonic mucosa. We will address this hypothesis in the following three well-integrated, but not mutually contingent specific aims, which will: (1) Define the role of NAD+ depletion vs. hyperPARylation as putative culprits in the pathogenesis of experimental colitis (2) Define the role of PARP1 and PARylation in colitis mediated by human IBD microbiota; and (3) Mechanistically define the roles of PARP1 in the highly interactive relationship between gut microbiota and epithelial barrier function and mucosal restitution.in the inflamed colon. This novel research plan will greatly advance the field of fundamental mucosal biology and will offer targeting of PARP1 activity as a potential IBD therapy.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY/ABSTRACT The University of Arizona Cancer Center (UACC) T32 Cancer Prevention and Control Training Program Addressing Health Disparities is designed to advance the independent careers of diverse scientists. The program benefits from the success of training nine postdoctoral Fellows during the first T32 grant period, all of whom are from underserved populations (including 5 Hispanic/Latino and 2 Native American scientists), access to the diverse population in Arizona who suffer significant cancer disparities, and on the 23-year history of UACC preparing outstanding cancer prevention and control (CPC) scientists. This T32 Program will support the training of four postdoctoral positions annually with a maximum of 10 postdoctoral trainees supported over five years. The program’s primary goal is to train a diverse workforce of independent CPC scientists working to reduce cancer health disparities. Our aims include: 1) Recruit outstanding and diverse applicants committed to careers in cancer prevention and control research with an emphasis on cancer health disparities; 2) Engage Fellows in a high quality, comprehensive training program that establishes their independence as cancer prevention and control scientists; and 3) Evaluate the program regularly and actively respond to program enhancement opportunities. The Program includes a structured curriculum in CPC and health disparities sciences while supporting individualized career development plans. The training program design supports trainee outcomes including independence in research and high productivity in academically relevant metrics of success (grants, manuscripts and scientific presentations). The Program is based on strong mentorship from a faculty representing diverse disciplines and deep experience working in diverse populations. Program evaluation is operationalized throughout the training to assure ongoing quality improvement. Specific fellowship activities of the training plan include: 1) Completion of required elements of the core curriculum and elective curriculum elements tailored to meet each postdoctoral Fellow’s training and professional goals; 2) Participation in mentored research experiences; 3) Submission and publication of peer-reviewed journal articles; 4) Presentation of work at scientific conferences; and 5) Preparation and submission of a research proposal for funding. This proposed training program aligns with the University of Arizona’s strategic plan and addresses the needs of our State’s unique population, including high numbers of Hispanic and American Indian residents, rural and border communities, aging population, and areas of persisting poverty. This distinctive geographic location along with our strong community ties, university partnerships, CPC postdoctoral training history, and university infrastructure and resources uniquely position us to continue to expand the next generation of diverse CPC researchers focused on reducing health disparities and working with underserved communities.

NIH Research Projects · FY 2024 · 2023-06

PROJECT SUMMARY/ABSTRACT We propose a better way to diagnose pulmonary embolism (PE) early and save lives. More than 900,000 people in the United States suffer from acute PE, and about 100,000 die each year. With 10% of such cases being fatal within the first hour of the onset of symptoms, rapid diagnosis of PE is critical to direct appropriate therapy. Unfortunately, clinical evaluation alone is unreliable and often results in grave diagnostic delays. Furthermore, while echocardiography at the patient’s bedside can rapidly detect heart dysfunction caused by PE, traditional echocardiography performed by cardiology services is not readily available in acute care settings. Thus, there is a critical need for use of a rapid, non- invasive diagnostic tool at the point-of-care (POC) to accurately assess for PE and direct emergency therapy. The focus of this research is to develop innovative artificial intelligence algorithms that can transform the care of patients with PE by enabling non-experts to use echocardiography to detect PE, direct emergency therapy, and improve survival. The rationale underlying this proposal is that the proposed artificial intelligence technology tools will provide a relatively simple and time-efficient strategy that can be implemented in most healthcare settings. This will, in turn, fulfill the overall goal of creating a positive shift in the management of patients presenting with PE. The proposed specialized artificial intelligence technology would ultimately be applicable to early detection of a wide variety of diseases. The long-term goal of our research is to develop and implement effective automated ultrasound tools that would significantly impact the diagnosis and treatment of different life-threatening conditions. The objective of this proposal is to develop and validate a prototype mobile artificial intelligence enabled-software platform that can accurately detect echocardiographic signs of PE. The hypothesis is that artificial intelligence algorithms will achieve levels of diagnostic accuracy equivalent to expert physician sonographers in detecting PE. This hypothesis will be tested by pursuing two specific aims: 1) Develop a machine learning algorithm for the detection of PE that can be extended to detect other cardiopulmonary conditions using explicit echocardiographic signs of PE and implicit image content representations. 2) Validate the accuracy of the machine learning algorithm to detect PE on echocardiographic images using explicit sonographic signs. Innovative reinforcement learning techniques will be utilized to accomplish the specific aims. The proposed research is significant because it will transform the care of patients with PE by enabling non-experts to use POC echocardiography. It will also have an immediate, positive impact because it will help lower morbidity, mortality, improve quality of life, and decrease healthcare costs by expediting diagnosis and therapeutic interventions. The proximate expected outcome of this work is improvement in the evaluation of patients with life-threatening PE by inexperienced healthcare providers, which will result in more accurate and rapid identification of cases that require emergency treatment. Our proposal aligns with the NIBIB’s overall mission to advance healthcare through innovative engineering and, more specifically, its emphasis on development of transformative unsupervised and semi-supervised machine learning technologies to enhance analysis of complex medical images and data for diagnosing and treating a wide range of diseases and health conditions.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY The University of Arizona (UArizona) MARC program provides research experiences, formal and informal mentoring, financial support and professional development training to upper-division underrepresented students who have an interest in and potential to pursue biomedical research careers. The program seeks to increase the number of such students entering PhD programs. The overarching objectives are to support 14 MARC trainees per year in two cohorts beginning June 2023 and for 85% of the trainees to complete the program, graduate from UArizona and matriculate in PhD or combined-PhD/MD programs by Fall 2029. The trainees will be underrepresented honors students from 14 biomedically-relevant majors distributed in 12 departments across four colleges at UArizona. Outstanding, well-funded UArizona MARC training faculty will provide research guidance and intensive mentoring, drawing upon their significant experience with undergraduates and their commitment to training underrepresented students. Trainees will also be formally mentored by the Program Director and Associate Director. Through the Minority Biomedical Research Colloquium series, trainees will meet prominent scientists from other institutions, as well as former UArizona MARC trainees, who are currently in or recently graduated from PhD programs. Trainees will attend workshops on responsible conduct of research, research safety and minority health disparities; take part in a scientific writing course; attend national scientific meetings; participate in intramural and extramural research; present posters at campus and national conferences; give oral presentations in the Research Colloquium series and other forums and attend workshops on the graduate application process. Each trainee will be provided individualized support on graduate school applications and will be coached through the interview and selection process. The UArizona MARC program will also encourage lower-division underrepresented students to perform well academically so as to secure MARC traineeships and thus prepare for careers in biomedical research. The MARC program has been iteratively designed based on feedback from former and current trainees, the program evaluator and the internal Advisory Committee. The success of this responsive program design is evident in metrics from the past funding cycle in that the percentage of underrepresented trainees entering biomedical doctoral research programs improved from 70% for the first cohort (2017–2018) to 90% for the fourth cohort (2020–2021). The MARC Program Director, Associate Director and Assistant Director have and will continue to provide persistent, seamless and inclusive support for trainees. We will continue to provide effective activities and inclusive support to enhance the transition of our trainees into biomedical research doctoral programs.

NIH Research Projects · FY 2025 · 2023-06

Abstract This grant proposal from the University of Arizona, Arizona Veterinary Diagnostic Laboratory is in response to Funding Opportunity Announcement (FOA) # PAR-22-063 from the Food and Drug Administration’s (FDA) Center for Veterinary Medicine (CVM) Office of Research Veterinary Laboratory Investigation and Response Network (Vet-LIRN). The purpose of this proposal from University of Arizona, Arizona Veterinary Diagnostic Laboratory (AzVDL) is to address the following three key project areas: 1. Participation in FDA/Vet-LIRN surveillance assignments and sample analysis in the event of animal food or drug-related illnesses or other large-scale animal food/drug emergency events requiring surge capacity testing of implicated diagnostic or animal food samples. 2. Providing analytical data using standardized methods, equipment platforms, and reporting methods; participation in proficiency testing and method training provided by the Vet-LIRN Program Office (VPO) at the Center for Veterinary Medicine; and implementation of standardized quality management systems for laboratories. 3. Participation in small-scale method development, method validation and/or matrix extension work as determined by the VPO. A successful implementation of the project outlined in the proposal is predicted for many reasons including the extensive experience of AzVDL’s professional and technical staff. The qualifications of the technical and professional staff are presented. As part of the proposal, a detailed view of the laboratory facilities and quality system are provided. This grant proposal also addresses the CVM request for sample analysis commitment as well as the laboratory’s ability and willingness to complete the proposed research in a timely manner. Several additional documents including the internal Proficiency Testing (PT) results (see Research Strategy), and certificate of AAVLD accreditation (appendix of Research Plan) are provided along with the

NIH Research Projects · FY 2025 · 2023-05

PERIPHERAL INFLAMMATION AND STRESS DRIVE VENTRAL STRIATAL MALADAPTATIONS PROJECT SUMMARY Mental illnesses such as depression and anxiety are a major disease burden linked to suicide and mortality. It is well known that exposure to stress can precipitate neuropsychiatric complications. The immune system also has a large influence on psychological symptoms and chronic conditions like inflammatory bowel disease dramatically increase risk of depression and anxiety. Surprisingly, little is known of the brain circuitry-specific mechanisms that drive this comorbidity. Our goal is to address this need by studying how systemic inflammation and stress cause maladaptations in reward/aversion circuitry of the ventral striatum (nucleus accumbens, NAc). In preliminary studies, we found that gastrointestinal (GI) inflammation, as a pervasive form of systemic inflammation, modulates stress-response behavior, and dysregulates NAc synaptic plasticity and excitability of D1 dopamine (DA) receptor (D1R) expressing medium spiny neurons (MSNs). Multi- omic exploration of the mechanistic basis for these effects implicated a novel dynorphin (DYN)-kappa opioid receptor (kOR)-Cdk5/p35-b adducin (ADD2) signaling cascade in the NAc which we hypothesize mediates these maladaptations. Based on these findings we propose to study the effects of peripheral inflammation, stress, and their interactions on neurobehavioral functions (Aim 1), and NAc synaptic plasticity, cell type-specific excitability, and DA neurotransmission (Aim 2). The novel kOR-Cdk5/p35-ADD2 pathway we have identified provides a mechanism by which maladaptive changes in DA neurotransmission can actuate alterations in DA-cAMP-PKA signaling and alter structural plasticity. We will study the mechanisms by which this pathway functions and its contribution to the effects of inflammation and stress on structural plasticity (Aim 3). Innovative components of this proposal include the study of inflammation/stress interactions, NAc cell type-specific interrogation of the role of kOR-Cdk5/p35-ADD2 signaling in mediating these effects, in vivo fiber photometry to study DA dynamics, and a novel systemic Cdk5 inhibitor as a targeted therapeutic approach. This research connects a strong field of striatal signal transduction to a major clinical problem. The impact will be to provide a detailed picture of the mechanistic basis for systemic inflammation-mental illness comorbidity and possible new approaches for therapeutic intervention.

NIH Research Projects · FY 2026 · 2023-05

Project Summary/Abstract The growing abundance of population genomic data creates a critical need for inference approaches that can reveal evolutionary history. The PI's long-term goal is to understand how natural selection shapes the evolution and function of the molecular networks that comprise life. Toward that goal, the PI's group develops and applies methods for inferring the evolutionary past from population genomic data. The objectives of this application are to understand how context affects mutation fitness effects, to develop improved inference methods, and to support the population genomics research community. The rationale is that this research program will both reveal new insights into evolution and enhance the ability of colleagues to reveal complementary insights. The PI's research group has expanded the concept of a distribution of fitness effects to multiple dimensions, focusing on differences in mutation fitness effects among populations. The PI proposes to apply this approach to numerous systems, to elucidate the relative roles of genetic and environmental context in creating differences in fitness effects. The group will also extend this approach to consider differences in fitness effects over time. The PI developed and maintains the software dadi, among the most popular approaches for fitting population genomic models to data. The PI will continue to support and enhance dadi, while developing complementary inference approaches. These will include new diffusion methods based on pairs of loci and the linkage among them and a novel deep learning approach for inferring the distribution of fitness effects. The PI helped found the PopSim consortium, which aims to expand the rigor and transparency of population ge- nomic models for the scientific community. The PI's group will continue to be active in the consortium, particularly leading a new initiative to facilitate rigorous testing of population genomic methods via open competition. The proposed research program is innovative both conceptually and methodologically. The novel concept of a multidimensional distribution of fitness effects has many applications, and the group will develop novel method- ology for several population genomics inferences. The expected outcomes of the proposed research are new insights into the ecology and biology of mutation fitness effects, new population genomic inference tools, and a framework for blinded evaluation of such tools. These outcomes are expected to have important positive impact on the filed of population genomics. The methods will be widely applicable and well-supported, and the inferences will feed into approaches for inferring the evolutionary past and predicting the evolutionary future.

NIH Research Projects · FY 2025 · 2023-05

ABSTRACT The current proposal requests an administrative supplement via NOT-OD-23-032 to support research continuity on NIH/NIDA R21DA058364 during a period of parental leave for the study MPI (Linde-Krieger). The goal of the parent study is to understand the role of social connectedness in OUD-related recovery outcomes, specifically during the postpartum fourth trimester, an ideal inflection point with untapped potential. The prevalence of opioid use disorder (OUD) during pregnancy has increased by more than 500% over the past 15 years. While motivation for and compliance with OUD treatment during pregnancy is heightened, up to 80% of postpartum individuals with OUD relapse to illicit opioid use within six months of childbirth. A growing body of evidence suggests that positive social connectedness and strong social bonds are associated with improved OUD recovery outcomes (e.g., reduced craving, lower risk of relapse). Conversely, loneliness and social isolation are significant predictors of opioid misuse and relapse, particularly for women. Loneliness increases during transitional periods including from pregnancy to postpartum, signaling increased risk for adverse postpartum recovery outcomes. To achieve this goal, we are actively pursuing three aims in the parent study. First, we evaluate differing theoretical models of social connectedness in this population by testing the main and stress-buffering effects of social connectedness on recovery outcomes up to one year postpartum (Aim 1). Additionally, we characterize dynamic changes in social connectedness across the fourth trimester and how patterns/changes relate to mothers’ recovery outcomes (Aim 2). Lastly, we complete in-depth qualitative interviews with 30 participants from the target population to explore the feasibility, acceptability, and opportunities for intervention to enhance social connectedness to improve the treatment of OUD and prevent postpartum relapse (Aim 3). The results of this work will directly contribute to scientific knowledge on the role of social connectedness in postpartum OUD recovery. Moreover, findings will identify new intervention targets, which will contribute to the development of novel, high-impact relapse prevention treatments tailored to the fourth trimester. Supporting the progress and continuity of this work through an NIH administrative supplement will be directly impactful to the 80,000-120,000 women, infants, and families who suffer the consequences of perinatal OUD every year. The requested supplemental funds will be of high value during a pivotal time in Dr. Linde-Krieger’s early career and will ensure continued progress and momentum on the parent award. Further, this supplemental funding will support the preparation a new NIH R-series grant submission by Dr. Linde-Krieger upon her return from parental leave.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Malaria parasites require pantothenate (Pan) from both the insect and mammalian hosts to synthesize coenzyme A (CoA) and acetyl-CoA (AC). Further, mosquito-stage parasites cannot take up preformed CoA from the insect host, so they are entirely dependent on mosquito Pan availability. Thus, we hypothesize that reducing Pan stores in the mosquito by increasing Pan kinase (PanK) activity and, in turn, CoA biosynthesis will limit parasite survival in the mosquito, without impacting the availability of CoA/AC to the mosquito itself. PanK is the rate-limiting enzyme in the CoA biosynthesis pathway and a logical target for our approach. In this study we will focus on increasing PanK activity in the mosquito to convert Pan into CoA and starve the malaria parasite of this essential precursor. To accomplish this we will utilize PanK-targeted small molecules or pantazines and genetic manipulation of PanK in our study host Anopheles stephensi. We will screen pantazines from a library of compounds developed by our collaborators at St. Jude Children’s Hospital. In Aim 1, we will use a Go-No Go strategy for pantazine screening that culminates in testing the capacity of selected pantazines to reduce P. falciparum and Plasmodium yoelii infections in A. stephensi. Aim 2 will validate the bioactivity and specificity of candidate pantazines identified in the screen in Aim 1. The specificity of candidate pantazines to activate PanK will be assessed through RNAi or CRISPR/Cas9 knockdown of PanK, followed by a characterization of the impact on Pan, CoA, AC and parasite infection success. Concurrent with Aims 1 and 2, we will generate transgenic A. stephensi with increased PanK activity and determine the impact on Pan levels and parasite survival in Aim 3. The generation of transgenic mosquitoes with increased midgut PanK expression will contribute to our assessment of PanK-dependent depletion of Pan stores on parasite infection as well as other aspects of mosquito biology related to vectorial capacity. Specifically, we will define the effects of mosquito PanK activation, via both pantazine treatment and molecular manipulations, on A. stephensi lifespan, stress responses, metabolism and reproduction. These studies will reveal important new insights into nutrient-driven mosquito-parasite interactions that drive parasite infection success and they will support future efforts to optimize pantazines and novel transgenic lines as distinct strategies for mosquito-targeted malaria control.

NIH Research Projects · FY 2026 · 2023-04

Abstract: Flaviviruses are primarily insect-borne, associated with global morbidity and mortality, and found on every inhabited continent. Unfortunately, current therapeutic options for treating diseases associated with these viruses are limited. All flaviviruses encode methyltransferases (MTases)—flaviviral NS5 for both N-7 and 2'-O methylations of viral genomic RNA. The N-7 MTase function is essential for replication of the viral RNA genome, whereas 2'-O MTase function is required for the virus to evade the host innate immune response. These activities are conserved among the flaviviruses. For this project, our collaborative team will optimize the current lead compounds, perform high throughput screening (HTS) to identify additional lead candidates, chemically optimize the lead candidates, and define structure activity relationships. Optimizing current lead compounds using cutting- edge medicinal chemistry, the team will perform a large scale HTS campaign using innovative fluorescence chemical probes to identify additional small molecule inhibitors of flavivirus RNA capping MTases. We will perform an in-depth investigation of the model of action and antiviral efficacy using in vitro biochemistry, structural biology, virology, in vivo pharmacokinetics, and in vivo animal models, which will allow the development of novel, effective, broad-spectrum, and druglike therapeutic agents against both flaviviruses. Preliminary progress has been made in the identification of initial lead inhibitors of these MTases, demonstrating low nanomolar antiviral activity. We will advance these compounds to further develop potent antiviral compounds while conducting large- scale screening in parallel for additional structural scaffold discoveries. Complementary expertise among our investigators will synergize and expedite the progress of this research. Our collaborative objective is to provide first-in-class drug candidates for the treatment or prevention of these viral infections.

NIH Research Projects · FY 2026 · 2023-04

Summary: The ability of an organism to reduce the brain blood flow in response to sudden surges in systemic blood pressure (BP) is known as cerebral autoregulation (CAR). In contrast to term neonates, preterm neonates are not able to reduce cerebral blood flow (CBF) in response to increased systemic BP. In preterm neonates, this exposes fragile cerebral vessels to a significantly increased blood flow at high pressure, leading to their rupture and brain damage. Our preliminary studies demonstrate that near-term fetuses can constrict carotid arteries and reduce CBF when systemic BP rises; however, this capability is not developed in the preterm fetus. We also observed that the constriction of carotid arteries to reduce CBF is regulated by the adrenergic nervous system, specifically by the activities of alpha-1 adrenergic receptors (α1-ARs). These receptors are expressed at a significantly lower number in preterm carotid arteries. Also, we observed that following the removal of adrenergic control in the near-term fetus by severing the superior cervical ganglion (SCG) made them lose their ability to reduce carotid blood flow (CaBF) to the brain with the rise in systemic BP. Thus, after the removal of SCG, both preterm and near-term fetuses cannot reduce CBF following an increase in systemic BP. During ex-vivo experiments on carotid segments, we observed that preterm arterial constriction in response to α1-ARs agonist was significantly lower than those from near-term lambs. Thus, we concluded that reduced activities of α1-ARs play a fundamental role in regulating CaBF with the rise in systemic BP. We also observed that the reduction in the activities of α1-ARs agonists in preterm resulted from reduced expression of α1-ARs compared to those in near-term fetal lambs. Furthermore, we present evidence that DNA hypermethylation reduces α1-ARs promoter activities by luciferase reporter assays and the involvement of histone modifications. Thus, we will test the hypothesis that promoter DNA hypermethylation and histone modifications reduce the expression and function of α1-AR subtypes (α1A-, α1B-, α1D) in the carotid arteries and play an essential role in the maturation of cerebral autoregulation from preterm to term fetus. We will also collect data from both sexes (male versus female) to identify sex-related changes. The studies will be conducted ex-vivo on isolated carotid arteries and in vivo in chronically catheterized fetal sheep. The hypothesis will be tested with two specific aims. Aim 1: From preterm to term fetus in a sex- specific manner, we will conduct an in-depth mechanistic analysis of promoter DNA methylation and histone modifications on differential expression of α1-AR subtypes in carotid arteries. Aim 2: From preterm to term fetus, in a sex-specific manner, we will determine the functional significance of differential α1-AR subtypes promoter methylation, histone modifications, and gene expression on carotid artery contractility and blood flow regulation to the brain in response to an increase in systemic pressure. The measurements will be conducted in real-time, in-vivo, with in-utero fetal maturation every week from 105 to 137 days. This will provide valuable information regarding the role of α1-AR subtypes and the epigenetic mechanisms involved in the maturation of CAR.

NIH Research Projects · FY 2025 · 2023-04

Heart failure progression is a complex biological process that is precipitated by the maladaptive myocardial response to injury, compounded by failure of the adult heart to replace lost or damaged cardiomyocytes. Conceivably, identifying common pathways that regulate these two seemingly unrelated processes would profoundly impact therapeutic strategies to prevent, and even reverse heart failure progression. Numerous observations by members of the proposed consortium and others support the notion that the endogenous capacity of the neonatal mammalian heart to proliferate fades in the early postnatal life as a switch from hyperplastic to hypertrophic growth of cardiomyocytes takes place. Members of the proposed consortium and others have also previously demonstrated that mechanisms linked to activation of immune response may play a role in cardiomyocyte hypertrophy, death, healing and even stimulation of new cardiomyocyte generation. The current proposal brings together several groups with significant expertise in immunology, myocardial remodeling, hypertrophy and regeneration with the overall goal of determining the role of immune response signaling in regulation of cardiac growth, healing and regeneration. Indeed, the immune response has taken center stage in the past several years as a primary determinant of both healthy cardiac aging and healing after injury, as well as a determinant of chronic disease states when it is inappropriately regulated. Thus, this Program Project will investigate a frontier and emerging area of scientific investigation involving the intersection between the immune system and the myocardium. Specifically, studies will examine the role of innate immune response signals in cardiomyocyte cell cycle regulation, as well as in myocardial remodeling and hypertrophy. In addition, dedicated studies will examine the role of cGAS-STING expression in cardiomyocytes and in immune cells in regulation of cardiac growth and regeneration. Finally, studies in the proposed Program Project will examine the role of macrophage subtypes in mediating myocardial response to cell therapy as well as injury. The proposed Program Project is well in line with the interest and expertise of the groups in the network and extends their focus into a highly relevant and poorly explored area of cardiac pathophysiology. The current proposal creates a new shared focus for all the groups in the Program Project that aims to establish a new paradigm in the field.

NIH Research Projects · FY 2024 · 2023-04

Project Summary/Abstract In the United States, millions of people sustain a traumatic brain injury (TBI) each year carrying individual, healthcare, and societal costs greater than $45 billion annually. Mild TBI (mTBI) accounts for more than 75% of all TBIs, with many individuals sustaining more than one. mTBI patients report post-concussion syndrome (PCS) symptoms that include sleep disorders (insomnia, daytime sleepiness), somatic symptoms (dizziness, headache, blurred vision), cognitive complaints (memory, executive function), and emotional problems (anxiety, depression, irritability, disinhibition). For many, PCS is transient, and still 10-25% report persistent PCS symptoms. The enigmatic PCS symptom presentation and persistence after mTBI urges investigation into dynamic responses in the brain that tie acute neurophysiology to behavioral function. The investigative team has refined in vivo imaging methods of miniaturized microscopes (miniscopes) to evaluate cerebral blood flow (CBF) and blood brain barrier (BBB) permeability in the unrestrained, behaving mouse. New preliminary data leverage the miniscope headcap to induce impact acceleration (weight drop) closed head injury. The data driving this proposal show immediate accumulation of fluorescent dextrans in the parenchyma within the field of view. The strength of this approach eliminates anesthesia during long-duration imaging, permits naturalistic behavior without head restraint, and locks into a baseplate for repeated imaging of single channel, wide-field, fluorescence. For the first time, the cumulative effects of TBI on neurophysiology (CBF, BBB permeability, sleep) can be regressed toward neurobehavioral impairments. The present proposal tests the hypothesis that the cumulative effects of mTBI on CBF and BBB permeability promote post-traumatic sleep and predict neurological impairments. Male and female mice are prepared for miniscope imaging through a cranial window and attached baseplate. The baseplate headcap substitutes for a helmet in closed-head impact acceleration TBI. With a 15 sec transition, miniscopes visualize CBF and BBB permeability with intraperitoneal dextrans (40-2000 kDa) in vasculature and parenchyma, respectively. The cumulative effects of mTBI are assessed with impacts delivered twice daily, daily, or every other day for a week. Aim 1 monitors the cumulative effects of mTBI with varying temporal spacing on CBF concurrent with post-traumatic sleep and subacute behavioral performance. Aim 2 quantifies the extent of dextran extravasation with post-traumatic sleep and subacute neurological performance. Neurological outcomes include anxiety (open field), spatial memory (novel object location), and somatosensory pain (mechanical hyperalgesia). Twice daily injuries likely show temporal summation of CBF, BBB permeability, and sleep effects, which are recovered with longer recovery times between injuries. The integration of miniscope imaging and closed head injury can propel future studies on physiological perturbations and clinical management of TBI.

NIH Research Projects · FY 2026 · 2023-04

Abstract The overall goal of this proposal is to determine the role of glyoxalase 1 (GLO1) in the pathogenesis of obesity, Type 2 diabetes (T2D), and non-alcoholic fatty liver disease (NAFLD). Greater than 34 million Americans have diabetes, and another 88 million are considered pre-diabetic. This is largely attributed to the prevalence of obesity, with 72% of American adults currently classified as overweight or obese. Among the comorbidities associated with T2D, over 70% of patients have NAFLD. Epidemiological studies have linked high fructose consumption with obesity, T2D, and NAFLD. This proposal identifies GLO1 as pro-NAFLD/obesogenic gene. Using CRISPR-Cas9, we have generated GLO1 knockout mice. When fed a high-fat high sucrose diet, these mice display significantly blunted weight gain, restored glucose tolerance, and reduced hepatic steatosis compared to wild-type counterparts. GLO1 is a ubiquitously expressed enzyme that detoxifies the glycolytic by- product, methylglyoxal (MGO). When GLO1 activity is disrupted, MGO levels increase, resulting in long-lived protein post-translational modifications. We have shown that MGO serves as a metabolic sensor for nutrient flux, regulating glycolytic output and transcriptional responses to sugar. Thus, we hypothesize that GLO1 is a pro- NAFLD/obesogenic gene, reducing MGO and removing the brakes on metabolism. We will test this hypothesis by addressing the following three Specific Aims: In Specific Aim 1 we will quantify the impact of Glo1 on whole- body energetics and hepatic lipid metabolism using a 16-week chow- or high-fat high-sucrose diet. We will quantify lean vs. fat mass, energy expenditure, and total activity. Hepatic fatty acid oxidation and mitochondrial respiration will be quantified in primary hepatocytes. Lastly, lipogeneic genes will be assessed using RNA-seq. In Specific Aim 2 we will determine the impact of Glo1 on intestinal fructose metabolism. The intestine is a primary site of fructose metabolism. Stable isotope labeling via 13C6-fructose oral gavage will be used to quantify intestinal, hepatic, muscle, adipose, and circulating fructose metabolites. This approach will quantitatively determine how Glo1 regulates intestinal carbohydrate metabolism, a previously unexplored area of research. Finally, in Specific Aim 3, we will confirm that MGO-derived histone PTMs regulate transcriptional responses to metabolism in vivo. Site-specific canonical and MGO-derived PTMs will be quantified in each tissue/cohort. Putative reader domains will be identified for MG-H1 histone modifications. Lastly, liver, intestine, muscle, adipose, and pancreas will be subjected to RNA-seq and ATAC-seq. This approach will determine the global epigenomic landscape across multiple tissues in a physiologically relevant model for obesity, T2D and NAFLD. Collectively, this proposal will combine mechanistic biochemistry with a multi-omics approach to determine the mechanisms by which GLO1 propagates disease progression.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract While immune checkpoint inhibitors (ICIs) have transformed the landscape of cancer treatment paradigm, the response rate is limited to a small subset of cancer patients (~20%). For colorectal cancer (CRC), the second leading cause of cancer-related deaths in US, only patients (~4%) with mismatch-repair-deficient or microsatellite instability-high tumors can respond to ICIs, leaving the vast majority of CRC patients with limited to no clinical benefit. Chemotherapy has been increasingly manifested to contribute significantly to the overall antitumor efficacy when combined with ICIs via switching the tumors from “immune-cold” to ‘immune-hot’. However, owing to the poor solubility and pharmacokinetics, limited tumor accumulation, and non-specific toxicities to healthy tissues, the utility of chemotherapeutics in enhancing the efficacy of ICIs has been considerably hindered. To render a safer and more efficacious chemotherapy-enabled immune response to cooperate with ICIs, our long- term goal is to develop an innovative and multifunctional liposomal nanotherapeutic platform via conjugating anticancer agents to the backbone phospholipid of liposome. We have developed a phospholipid-derived camptothecin (CPT) liposome (Camptothesome) nanoplatform, which significantly prolonged blood circulation time, enhanced tumor uptake and therapeutic efficacy and minimized systemic toxicities compared to free CPT. Moreover, Camptothesome potentiated the anti-CRC efficacy of PD-L1/PD-1 inhibitors, resulting in partial eradication of tumors in immunocompetent mice. To improve the efficacy of this combined therapy, we used Camptothesome to co-deliver an inhibitor targeting another independent immune checkpoint, Indoleamine 2,3- dioxygenase (IDO1), which markedly enhanced anti-CRC efficacy and immunity. To further strengthen the delivery efficiency and explore the potential of this nanoplatform in enhancing PD-L1/PD-1 blockade therapy, in this proposal we will: Aim 1: Improve the Camptothesome system for enhanced therapeutic delivery. Aim 2: Determine the tumor delivery efficiency and pharmacokinetics of the improved co-delivery system in murine CRC models. Aim 3: Define antitumor effects of the improved co-delivery system with or without PD-L1/PD-1 blockade in murine CRC models. The mechanistic action for the in vivo antitumor efficacy and immune responses of the combined therapy will also be elucidated. Successful completion of this proposal will result in an innovative and multifunctional nanotherapeutic platform for improved and safe CRC immunochemotherapy. Moreover, given that IDO1 is expressed in diverse cancer cells, and the broad applicability of this nanoplatform to other anticancer drugs, our combination nanotherapeutic system has the potential to revolutionize cancer treatment paradigms.

NIH Research Projects · FY 2026 · 2023-04

SUMMARY: Exercise as an Immune Adjuvant for gd T-cell Therapies in Hematologic Malignancies gd T-cells are being considered as an alternative to standard CAR ab T-cells for treating leukemic relapse after hematopoietic stem cell transplantation (HSCT), largely due to their ability to function across MHC barriers without causing graft-versus-host disease (GvHD)1. gd T-cells can be readily expanded in vitro and in vivo using zoledronate (ZOL) and have demonstrated anti-tumor activity in preclinical and early phase clinical trials, but their efficacy against CD19-expressing tumors including acute lymphoblastic leukemia (ALL) and non-Hodgkin’s lymphoma (NHL) has been modest3. Recently, CD19 CAR gd T-cells were found to have profound effects against CD19+ tumors in vitro and in xenogeneic mice, albeit inferior to CD19 CAR ab T-cells, although CD19 CAR gd T-cells were more effective at eliminating CD19 negative escape variants5, 6. As such, if the natural cytotoxicity of gd T-cells could be enhanced they would become a highly attractive “off the shelf” therapeutic option for ALL and NHL. Our goal is to improve gd T-cell therapeutics by collecting “superior” gd T-cells that have been mobilized to peripheral blood by exercise or a synthetic b2-adrenergic receptor (AR) agonist and arming them with a CAR. We will build on several novel and important observations we have made: (i) a single exercise bout instantaneously mobilizes gd T-cells bearing a cytotoxic, co-stimulatory and tissue migration phenotype, allowing their ex vivo manufacture with ZOL+IL-2 to increase by 100-300%4; (ii) exercise expanded gd T-cells have higher in vitro cytotoxicity against several hematologic tumors4 and are more capable of inhibiting K562 leukemic growth in xenogeneic mice, particularly when combined with ZOL sensitization; (iii) exercise skews expanded gd T-cells toward an activated phenotype with heightened NKG2D, TRAIL, DNAM-1 and lowered NKG2A expression, and blocking these activating receptors, or their ligands on K562 cells, abrogates the exercise effects on gd T-cell cytotoxicity; and (iv) the mobilization of these superior gd T-cells with exercise is driven by b2-AR activation4. We hypothesize that exercise will also enhance the quality of CAR gd T-cells by mobilizing gd T-cells with sustained activation of cytotoxicity, co-stimulation, oxidative phosphorylation, homing and proliferation related genes, and that this mobilization will be precipitated by increased cAMP signaling. Our aims are: 1) Determine if a single exercise bout can improve the quality of CAR gd T-cells expanded from healthy donors. 2) Explore the transcriptomic basis for the enhanced expansion and cytotoxicity of exercise mobilized gd T-cells and expanded products. 3) Identify the b2-AR signaling pathways responsible for mobilizing gd T-cells with enhanced expansion and cytotoxicity potential. Our approach involves the use flow cytometry, xenogeneic mouse models, single cell RNA sequencing, and comparisons with CD19 CAR ab T-cells in human trials involving exercise with b-blockers and b-agonist infusion models. We expect these aims to identify underpinning mechanisms and pave the way for a clinical trial whereby exercise/b-agonist mobilized gd T-cells can be collected from donors and cancer patients to increase the potency of CAR T-cell therapies to treat refractory disease and relapse after HSCT.

Fonds de recherche du Québec – Santé · FY 2023-2024 · 2023-04

Volet: Formation postdoctorale - Citoyens canadiens et résidents permanents; Domaine: Vieillissement; Objet: Maladies neurodégénératives; Objet: Histologie; Application: Santé; Application: Fondements biomédicaux de la santé humaine; Mots-clés: MALADIE DE PARKINSON, BIOMARQUEUR, HISTOLOGIE, BULBE OLFACTIF , OLFACTION, IMMUNOFLUORESCENCE

NIH Research Projects · FY 2026 · 2023-02

Heart failure is a devastating disease with mortality rates exceeding many malignancies. The pathophysiological basis of systolic heart failure lies in the inability of the adult mammalian heart to regenerate lost or damaged myocardium. Although limited cardiomyocyte turnover does in fact occur in the adult mammalian heart, it is insufficient for restoration of contractile function following injury. In contrast to the adult mammalian heart, my group has shown that newborn mammals have a remarkable endogenous myocardial regenerative capacity, mediated by proliferation of preexisting cardiomyocytes. Nevertheless, the mere realization that the heart is not a post-mitotic organ created a lot of excitement in the past two decades and led to a flurry of bench and clinical studies aimed at outlining the cardiac regenerative potential of various cell types. While many of these studies may hold therapeutic promise, mounting evidence suggest that cell therapy may enhance some endogenous repair or regenerative mechanisms such as stimulation of cardiomyocyte proliferation. Importantly, current evidence suggests that both the regenerative ability of the early postnatal heart, and cardiomyocyte turnover in the adult heart are mediated by proliferative competency of pre-existing cardiomyocytes. However, mechanisms of regulation of mammalian cardiomyocyte cell cycle arrest shortly after birth remain poorly understood. Therefore, we believe that a program focused on understanding mechanisms of cardiomyocyte cell cycle regulation could inform future therapeutic interventions for heart regeneration. Our studies are focused on three broad questions: 1) Is loss of the regenerative capacity of the mammalian myocardium an evolutionary tradeoff to gain metabolic efficiency? 2) How is the slow turnover of cardiomyocytes in the adult heart regulated? 3) Does cardiac mechanical load represent a regenerative block

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY/ABSTRACT A hypothesis of aging is that the accumulation of cellular damage can lead to tissue malfunction and organismal deterioration. A key mechanism for maintaining cellular homeostasis and preserving cell function is autophagy, a hydrolytic cellular recycling process whereby cytosolic materials, referred to as cargo, including lipid droplets (LDs) and damaged proteins, are degraded in the lysosome. In turn, aberrations in autophagy can result in the accumulation of different toxic cytosolic contents, which is a molecular signature of many age-related disorders, including neurodegeneration. While there is a prominent functional link between autophagy, aging and diseases, the molecular mechanisms that cause the age-dependent decreases in autophagy remain unclear. Notably, autophagy can also selectively recruit one type of molecule for degradation. Recent studies support the hypothesis that selective autophagy plays a crucial role in combating chronic diseases. Several human brain post-mortem studies have uncovered lipid species that accumulate in brains affected by Alzheimer’s disease (AD), possibly impeding neuronal function and thereby contributing to neurodegeneration. Therefore, discovering different interventions that can be used to affect lipophagy (LD turnover) selectively may be ideal for tackling lipidotoxicity-linked AD. However, such pharmacological or genetic tools are currently unavailable. Furthermore, selective cellular factors that can facilitate LD recruitment for lipophagy remain unknown. In this proposal, I aim to address these greater needs in understanding the regulatory mechanisms of lipophagy and its function relevant to aging and neurodegenerative disorders. Our lab recently performed a cellular LD clearance high-throughput screen to identify small molecules and pathways that induce selective lipid clearing autophagy for slowing age-related diseases. Among these, we identified compound A20 that clears lipids in an autophagy-dependent manner in the nematode C. elegans to promote healthspan and lifespan. Emerging evidence suggests that A20 may act via lipophagy to clear lipids. I hypothesize that uncovering the lipophagy mechanism utilized by A20 will help us identify novel lipophagy regulators. Furthermore, since lipid accumulation is now linked to AD, I will employ a novel human AD patient- derived organoid model (3D neuronal culture with astrocytes) to determine whether A20 normalizes the lipid- linked pathogenic signature and normalizes pathogenic molecular phenotypes. Finally, I will characterize the functional changes in lipophagy and lipid homeostasis during AD using these human-derived organoid models. My studies are significant, as they will help us generate new mechanistic insights towards lipophagy activation during aging linked to AD. Such knowledge is vital to further our understanding of diseases exhibiting a lipophagy deregulation component. Furthermore, completion of these studies may potentially reveal strategies that could be used to combat neurodegenerative diseases.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Overview Environmental Health Sciences Research for Indigenous Scholar Engagement (EHS-RISE) is a 5-year summer research training program for Native American undergraduates at the University of Arizona (UArizona). It is housed within the Undergraduate Biology Research Program, the Department of Pharmacology and Toxicology, and the Southwest Environmental Health Sciences Center. The long-term goal of this program is to increase the number of Native Americans in STEM fields. The short-term goal is to develop and retain a stable cadre of Native American students who graduate from University of Arizona with STEM degrees. These goals will be realized through the intentional construction of a research training environment that supports and integrates Native values for Native students pursuing a science degree. The specific objectives of this program are: Objective 1: Identify, recruit, and retain nine Native American undergraduate students annually for five years. Objective 2: Develop and expand trainees’ scientific skills and knowledge. Objective 3: Foster and sustain trainees’ level of engagement. Objective 4: Embed the program within a culturally relevant environment and learning community. These objectives will be achieved by addressing each of the eight pillars of the American Indian Well Being Model in Higher Education within the context of a rigorous environmental health research training program. Intellectual Merit EHS-RISE approaches STEM diversity from a non-traditional mindset. Instead of recruiting Native students into a program based on a Western scientific model, we have built a program that integrates Native values and builds bridges between Indigenous and Western science and knowledge. It is a model which equally values the contribution of both epistemologies. In this way, we are advancing STEM workforce diversity and inclusion by honoring and integrating the impact and influence of Native perspectives and values in science. Broader Impact A diverse workforce, in which the contributions of all members are valued, results in greater innovation, creativity, engagement, and decision-making. Our model supports the development of culturally confident and scientifically competent Native American scientists who will bring their values and knowledge into the STEM workforce, thus advancing science and innovation.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY/ABSTRACT Cerebral palsy (CP) is a major neurodevelopmental disorder affecting an estimated 1:323 individuals. CP affects the motor stream of development, and people with CP may have difficulty walking, talking, eating, or using their hands. Some may have involuntary muscle activation and/or movements that may be painful and/or disruptive, affecting quality of life and inclusion. Although environmentally-mediated causes of CP such as perinatal stroke, prematurity, prenatal infection or lack of oxygen have been known to lead to CP for some time, recent findings from our group and others indicates that for as many as one-third of individuals with CP, a genetic mutation may be the cause of their CP symptoms. Identifying the genetic basis of CP is important because our data suggests that the identification of a genetic cause of a patient’s CP can lead to a change in management > 20% of the time and this number is expected to grow steadily. Genetic forms of CP may occur sporadically in approximately one-half of cases. However, ~1/3 of genetic cases of CP are estimated to exhibit autosomal recessive inheritance, wherein both parents are unaffected carriers of the condition. Most current research in CP is focused on parent-child trio-based studies. We propose a different approach, focusing instead on families with multiple affected children with CP inter- related by blood. We have enrolled hundreds of such families in collaborations with physician colleagues in the Middle East, Africa and Southeast Asia. Studying highly genetically-informative families such as these is anticipated to represent a powerful approach for cerebral palsy-associated gene discovery. The signs and symptoms of the families we have enrolled in our studies have been diligently catalogued, including assessments by multiple specialists, and the collection of facial photographs and patient videos that highlight their movement disorders. These resources, as well as detailed laboratory and radiological assessments will allow us to comprehensively catalog the symptoms of the individuals we have partnered with for these studies. We will then perform whole exome sequencing to identify changes in the genome found in affected members of the family but not in healthy individuals. Once a candidate gene is identified, we will connect with colleagues worldwide to find patients with similar symptoms who harbor variant(s) in the same gene. We will then conduct a series of laboratory analyses in order to confirm or refute a role for the candidate gene in CP. This will include both classic and massively parallel unbiased studies of RNA, protein, and/or metabolites, and assessment in complementation studies, cell-based assays, fly models and human induced pluripotent stem cell-neurons/astrocytes from patient samples. Impact: We anticipate that this approach will allow us to identify dozens of novel CP-associated genes, which in turn will facilitate clinical advances in diagnosis and management as mechanism-based therapies continue to be developed.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT TGF-β affects virtually all aspects of mammalian physiology starting from early embryonic development to adult tissue homeostasis through regulation of diverse cellular functions including proliferation, differentiation, and apoptosis. TGF-β signaling also plays an important role in cell metabolism, although there has been incremental progress in understanding how it differentially regulates mitochondrial biogenesis, respiration, and organelle destruction. While such varying effects are theorized to occur primarily through slow-acting contextual gene regulation, TGF-β is also capable of inducing more rapid, direct, and reversible changes in mitochondrial shape and function through largely unknown mechanisms− a key aspect that represents an important knowledge gap in the field. Our research program has focused on two powerfully opposing mechanisms by which two major TGF-β effectors, Smad2 and TAK1, control mitochondrial fusion/fission dynamics to achieve and maintain metabolic homeostasis. We seek to understand mechanisms governing their organization, activation, and regulation in mitochondrial remodeling and how they influence cell behavior using angiogenesis as a developmental model system. Our studies will provide unique perspectives on how the complex TGF-β signaling networks control mitochondrial dynamics to affect their metabolic, developmental and homeostatic roles in vascular physiology.