University Of Alabama At Birmingham

NIH Research Projects · FY 2025 · 2021-06

Project Summary / Abstract Abnormal mucociliary clearance (MCC) is a critical component of cystic fibrosis (CF) lung disease, and is postulated to contribute to the high incidence of chronic pulmonary infections in this patient population; in turn the presence of chronic infection is thought to worsen the MCC defect, creating a cycle of mucus obstruction, infection, and inflammation that is difficult to interrupt or reverse. However, the mechanisms and interactions responsible for this phenomenon are not well understood. New animal models, such as the CF rat, developed at our institution, have been useful in identification of key factors that lead to chronic infection with the pathogen Pseudomonas aeruginosa in the CF airway. This animal model develops the MCC defect progressively, providing a model with which to study patients with early disease as well as late disease. In this model of CF, mucus must be abnormal before exposure to Pseudomonas aeruginosa to convert the infection to a chronic phenotype. CF rats exposed before the mucus abnormality develops are able to clear the infection. A new rat model harboring a humanized G551D-CFTR genomic insert respond to FDA-approved CFTR modulators that treat the fundamental defect of CF disease. Using the innovative Micro-Optical Coherence Tomography (µOCT), a high-resolution reflectance imaging modality that can simultaneously and non-invasively evaluate airway hydration, ciliary beating and mucus transport and viscosity in situ, we can analyze aspects of the mucus defect in both the CF rat model before and after infection, with or without CFTR modulators. Using these tools, this proposal will seek to investigate the mechanisms that cause patients with CF to transition acute infections into chronic ones, with the following independent but complimentary aims: 1. Determine if Muc5b is the specific component of mucus that promotes chronic Pseudomonas aeruginosa infection. 2. Determine if inflammation is necessary and sufficient to accelerate the mucus defect, predisposing the airway to chronic P. aeruginosa infection. 3. Determine if new highly effective CFTR modulators promote clearance of P. aeruginosa by normalizing abnormal mucus in the airway. This proposal will determine the early events that lead to infection and progression in CF pulmonary disease and how this relates to the conversion of P. aeruginosa from intermittent to chronic in this patient population, using a highly relevant animal model. The studies will provide new fundamental observations that will inform our understanding of the CF respiratory pathology and help identify robust therapeutic targets suitable for intervention.

NIH Research Projects · FY 2025 · 2021-06

Eukaryotic cells deploy at least three multisubunit, DNA-dependent RNA polymerases to express the robust variety of RNAs required for cell survival and proliferation. In contrast to prokaryotic cells which express a single RNA polymerase, eukaryotic RNA polymerases have evolved into specialized cellular roles. RNA polymerase I (Pol I) synthesizes the majority of ribosomal RNA, Pol II synthesizes messenger RNA and most regulatory RNAs, whereas Pol III synthesizes transfer RNA and the 5S ribosomal RNA. While this "division of labor" among the nuclear Pols has been appreciated for many years, substantial gaps in our understanding of the functional divergence between the eukaryotic RNA polymerases remain. How do the enzymatic properties of these enzymes differ? How do these functional differences influence regulation of transcription by associated transcription factors? How do divergent enzymatic properties of the Pols impact their unique cellular roles? The answers to these questions are fundamentally important for understanding eukaryotic biology. It is well established that the rate of ribosome synthesis is proportional to the rates of cell growth and proliferation. As a consequence, several labs around the world seek novel inhibitors of ribosome synthesis as potential anticancer chemotherapy agents. Since transcription of the ribosomal DNA by Pol I is the first, rate-limiting step in ribosome synthesis, Pol I has emerged as a key target for the development these inhibitors. In order to selectively inhibit one RNA polymerase without affecting the others, it is crucial to understand the fundamental properties of the enzymes as well as the cellular mechanisms by which the enzymes are controlled. Thus, defining Pol I activity and its cellular roles has immediate translational value. By defining the landscape of eukaryotic transcription and the many cellular roles of Pol I, this project will reveal answers to fundamental questions in evolutionary biology while informing ongoing studies aimed at therapeutic targeting of ribosomal RNA synthesis. To reach these goals, our lab has developed a robust platform of experimental approaches including biochemical, biophysical, genetic, and genomic methodologies. We have pioneered the development of both the experimental and analytical strategies required to rigorously and thoroughly define the functional properties of RNA polymerase I. The overall goal of this project is to leverage these experimental strengths to impact our overall understanding of eukaryotic gene expression and to lay the foundation for ongoing and future studies aimed at therapeutic inhibition of Pol I.

NIH Research Projects · FY 2025 · 2021-05

The goal of this investigation is to characterize the influence of the DNA methylome central to the increased risk of Monoclonal Gammopathy of Undetermined Significance (MGUS) observed in high-risk populations. MGUS precedes Multiple Myeloma (MM), which is characterized by cellular resistance to apoptosis leading to prolonged survival and accumulation of clonally expanded, cytogenetically heterogeneous, antibody producing tumor cells in the bone marrow and extramedullary sites. Beyond a few well-established risk factors, the etiology of MGUS/MM remains largely unknown. Although evidence suggests a germline component, inherited alterations in DNA sequence alone does not explain the risk. Advances in epigenomics offer new opportunities to characterize the heritable changes in gene activity, or plasticity in germline variation due to past exposures, which could significantly improve our understanding MGUS/MM etiology and provide new insight for improved clinical monitoring in high-risk populations. We will test the overarching hypothesis that distinct methylome signatures correlate with the excess risk of MGUS in high-risk populations by altering target gene expression. Using an epigenome-wide approach, we will capitalize on a unique opportunity to explore differentially methylated positions in DNA obtained from a network of well-characterized, treatment naïve populations of MGUS and MM while taking advantage of recent technological and analytic advances. This project leverages existing partnerships, resources and comprehensive, high-quality clinical data and biospecimens systematically collected in a well-characterized network of treatment-naïve populations, to improve our understanding of MGUS-MM etiology and to advance a set of biomarkers required to improve efforts to predict and manage MGUS-MM clinical course in high-risk populations.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY: Non-alcoholic fatty liver disease (NAFLD), once rarely observed in children and adolescents, is now the most common form of liver disease in this age group. If untreated, NAFLD can progress rapidly from simple fat infiltration to advanced stages of the disease including steatohepatitis, fibrosis, and cirrhosis ultimately requiring liver transplantation. It has been reported that approximately 20% of patients with NASH will progress to cirrhosis and liver failure. However, patients with NAFLD universally have hepatic insulin resistance placing them at high risk of developing glucose intolerance and type 2 diabetes (T2D). The pathogenesis of hepatic insulin resistance is closely tied to lipid accumulation, suggesting that depletion of hepatic lipid is critical in the prevention of T2D in pediatric populations. Currently, no pharmaceutical treatment exists to directly reverse NAFLD and limited progress has been made to identify effective, non-invasive treatment. Because lifestyle changes remain the mainstay therapy for children with NAFLD, there is urgent need for evidenced-based guidelines on the optimal dietary approaches to safely and effectively reverse disease course. If fatty liver and insulin resistance can be reversed during the developmental adolescent years, it may be possible to prevent progression of the disease to advanced stages and prevent occurrence of other diseases such as T2D and CVD. Our preliminary data in 23 adolescents with NAFLD suggest that a weight-maintaining, low glycemic, moderately CHO-restricted diet (~100 g CHO/day) significantly improves both hepatic steatosis and hepatic insulin sensitivity. After 8 weeks, we found the CHO-restricted diet resulted in significant reduction in hepatic lipid content (-6.0±4.7%, p<0.001), whereas no change in hepatic lipid was observed in the fat-restricted (control) diet group. HOMA-IR, an index of hepatic insulin resistance, was reduced (improved) (-1.2±5.1, <0.05) in the CHO-restricted group, and increased (worsened) slightly in the fat- restricted (control) diet group. While these results are encouraging, this study needs to be verified in a larger sample before translation to clinical practice can be recommended. It is also critical to measure hepatic insulin sensitivity using accepted, rigorous methods to determine if the reduction in hepatic lipid is associated with improvement in hepatic insulin sensitivity. Further, data identifying the changes in biological processes, such as de novo lipogenesis, β-oxidation and lipid metabolism, that lead to depletion in liver fat in response to a energy balanced CHO-restricted diet in the absence of significant weight loss in pediatric populations are needed. The proposed study will address these gaps in knowledge using a 6 month family-based intervention with 2 phases, a 12-week controlled feeding phase and a 12-week free living phase, to examine the effects of two weight maintaining diets (moderately CHO-restricted vs fat-restricted diet) on depletion of hepatic lipid content, improvement in hepatic insulin sensitivity, and changes in the plasma metabolome in adolescents with NAFLD.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY Randomized controlled trials by Basch (collaborator) and colleagues demonstrated that weekly electronic home-based PRO symptom monitoring with automated alerts to clinicians (Home ePRO) in cancer patients was associated with reduced healthcare utilization, improved quality of life, and increased overall survival. However, these trials were administered using infrastructure supported by research funding. A knowledge gap remains about optimal implementation strategies for and effectiveness of Home ePROs in real-world settings. To address this gap, investigators from two institutions will conduct a large-scale, population-based implementation of an evidence-based Home ePRO intervention for all adult cancer patients receiving chemotherapy, including vulnerable populations such as African Americans, rural residents, and socioeconomically disadvantaged individuals. This implementation will leverage infrastructure from Medicare's payment reform projects (Oncology Care Model, Oncology Care First Model), which require and financially support patient navigators, a natural workforce for Home ePRO implementation. Our hypothesis is that the deployed implementation strategies will result in successful navigator-delivered Home ePRO, which will improve both patient and health system outcomes. Using the Consolidated Framework for Implementation Research (CFIR), this hypothesis will be tested using a hybrid type 2 design with three specific aims: 1) evaluate implementation of navigator-delivered Home ePRO for all cancer patients across multiple practice sites; 2) examine the barriers, facilitators, and implementation strategies used in implementing navigator- delivered Home ePRO; and 3) assess the impact of Home ePRO on clinical and utilization outcomes. In Aim 1, Home ePRO will be evaluated using implementation outcomes (service penetrance, provider adoption/penetration, intervention fidelity). In Aim 2, we will gauge patient and healthcare team perceptions of barriers and facilitators of navigator-led Home ePRO, implementation strategies used to address these barriers, implementation strategy fidelity, and perception of implementation strategy ability to address barriers using an iterative qualitative analysis. In Aim 3, patient-level outcomes (functional status, distress, depression, healthcare utilization, cost, survival) will be evaluated using real-world data sources. The project is innovative because it will be the first study to evaluate real-world implementation of navigator-led Home ePRO for all cancer patients receiving chemotherapy, an approach that is both immediately scalable and sustainable within value-based payment models. The proposed research is significant because it is expected to demonstrate successful implementation of navigator-delivered Home ePRO and effectiveness of the ePRO intervention on diverse patient populations. Furthermore, the project will generate an implementation blueprint of successful implementation strategies that can be easily applied to other patient-reported outcomes, with the potential to positively impact patient care as health care transitions to a value-based system.

NIH Research Projects · FY 2025 · 2021-05

Project Abstract Approximately 80% of early stage head and neck tumors undergo surgery of curative intent at some point in their treatment course. The success of this oncologic surgery primarily depends on obtaining clear surgical margins. Despite best efforts to obtain wide margins, the surgeon always struggles with removal of healthy, functional tissue. This resulting tension often results in close (<5mm) or positive (tumor at cut surface) margins in 30% of head and neck cancer resections. We have developed a strategy that could improve both intraoperative and ex vivo margin assessment using a fluorescently labeled therapeutic antibody as a targeted imaging agent. We hypothesize that fluorescence imaging of a systemically injected fluorescent anti-epidermal growth factor receptor (EGFR) antibody (EGFR is over-expressed in 80-90% patients with head and neck cancer) will direct the attention of the surgeon and pathologist toward areas that are more likely to be tumor-positive and reduce sampling error. This will lead to improved identification of tumor-positive margins, and thus improved surgical outcome. 1

NIH Research Projects · FY 2025 · 2021-04

The purpose of this project is to determine if the relationship between a history of childhood maltreatment (CM) and suicide risk is associated with alterations in the expression and epigenetic modification of specific microRNAs (miRNAs), thereby providing a molecular signature of suicide risk in people with a history of CM. We propose that whereas both major depressive disorder (MDD) and suicidality are complex phenotypes, CM alters the risk threshold for both. Epigenetic changes caused by early stressful events can induce long-term alterations affecting networks of genes. miRNA expression represents one of the central mechanisms for environmental regulation of gene expression. miRNA sequences themselves are epigenetically modified. The sum of these effects may explain long-term cellular (mal)adaptations which may lead to suicide vulnerability in the CM population. Using a specific cell surface marker, we isolated neural-derived exosomes from blood plasma and found that these exosomes were not only enriched with brain expressed miRNAs, but also showed a unique set of miRNAs that were associated with CM and suicidality. Changes in the same set of miRNAs were also noted in the brain of suicide subjects with a history of CM. In addition, suicidal subjects with and without CM showed differential regulation of miRNAs in response to acute stress, a short-term risk for suicidal behavior, particulay in the context of CM. Moreover, miRNA expression changes were highly correlated with exosomal miRNA promoter methylation. Based on our preliminary data, we propose an overarching hypothesis that there are multiple paths to suicidal behavior, and CM represents a unique path that is associated with altered expression and epigenetic modification of a specific set of miRNAs and concomitant downstream specific target genes and network(s). To test this, we will: 1) identify a set of neural-derived exosomal miRNAs that are associated with the interaction of suicidality and CM severity while controlling for the independent effects of suicidality, CM, and MDD; 2) examine whether an acute experimental stressor differentially impacts the expression of neural-derived exosomal miRNAs in suicidal patients with and without CM; 3) use bioinformatic tools to examine potential mechanisms by which altered neural-derived exosomal miRNAs may contribute to CM-associated suicidal behavior; and 4) examine if changes in CM-associated miRNAs are explained by modifications in their DNA methylation. In participants (n=450; replicated in a cohort of 350 subjects) across the spectrum of MDD, suicide, and CM severity, we will test for the main effects of each, and identify a subset of miRNAs that are associated with suicidality and CM severity. We will also test for the independent main effects of CM or suicidality on miRNAs and the interactions among those factors. Altogether, using a distinct study population, unique neural-derived plasma exosomes, and innovative molecular, biostatistical, and bioinformatic tools, our study will identify: 1) neural-derived plasma exosomal miRNAs as a novel biosignature of suicide risk in the context of CM that can be tested in longitudinal studies, and 2) potential mechanisms by which CM can act as a risk factor for suicidality.

NIH Research Projects · FY 2025 · 2021-04

Project Summary We propose to investigate the contribution of the long non-coding RNA (lncRNA) Neat1 in astrocytes and identify the epigenetic mechanisms in these glia cells involved in enhancing memory resiliency with age. Interventions to enhance memory resilience within the aging population are possible. However, research studies that inform resiliency in this area are still lacking. Within the aging hippocampus, it is now clear that abnormal epigenetic control of gene transcription contributes to memory deficits. Nearly all the suggested epigenetic mechanisms controlling memory formation have mostly been attributed to neuronal cells within the hippocampus, largely disregarding these mechanisms in astrocytes. Thus, little is known about how astrocytic epigenetic mechanisms influence memory resiliency with age. Our long-term goal is to study the role of lncRNAs in astrocytes and to identify how these powerful epigenetic regulators impact memory formation with aging. Our pilot data demonstrate that Neat1 expression is decreased in area CA1 of young adult mice and overexpressed in aged mice. Furthermore, we demonstrate that inhibiting Neat1 expression in area CA1 of the hippocampus of aged mice reverses memory impairments. Pilot studies also suggest a strong relationship between G9a mediated H3K9me2 hypermethylation with Neat1 overexpression in aged adults. Based on these preliminary results, we plan to rigorously investigate the effects of manipulating Neat1 in astrocytes and determine effects on age-related memory decline. To gain further mechanistic insight into Neat1 mediated gene transcription in astrocytes in the aging hippocampus, we will use state-of-the-art approaches such as CRISPR reprogramming and chemogenetics to elucidate the epigenetic mechanisms in astrocytes in our aging animal model system. Our overarching hypothesis is that Neat1 contributes to age-associated changes in hippocampal astrocyte diversity, astrocyte function and vulnerability to memory dysfunction. Our Specific Aims are as follows: Specific Aim 1: Test the hypothesis that Neat1 is associated with astrocyte diversity with aging; Specific Aim 2: To determine the mechanism by which Neat1 acts to influence chromatin restructuring in astrocytes from young versus aged animals; and Specific Aim 3: To determine the contribution of astrocytic Neat1 to memory resiliency with age. Collectively, these studies will identify epigenetic mechanisms in astrocytes involved in age-related memory decline, with broad implications for treatment options for age-related dementia and Alzheimer’s disease.

NIH Research Projects · FY 2026 · 2021-04

PROJECT SUMMARY Long-acting (LA) antiretroviral (ARV) formulations represent major advancements for ending the HIV epidemic in the U.S., both on the treatment side, through antiretroviral therapy (ART), and prevention side, through pre-exposure prophylaxis (PrEP). However, major gaps in viral suppression and prevention still remain, particularly in the U.S. South and among subpopulations such as adolescents and young adults (AYA, ages 15-24). AYA vulnerable to HIV face unique challenges in persistence to ART or PrEP, and LA ARVs have the potential to help overcome some of these challenges through greater confidentiality and reduced stigma compared to oral ARVs. LA ARVs are now entering routine clinical care in the U.S., yet implementation evidence gaps persist on their scale up among AYA in U.S. South; these gaps can be filled by lessons from countries, such as Kenya and Botswana, rapidly scaling up LA ARVs. By directly comparing implementation experiences across U.S. and international settings, this study will generate actionable evidence to inform U.S. HIV treatment and prevention strategies, supporting NIH priorities to optimize delivery of LA ARVs for AYA. Another major concern for AYA is sexual and reproductive health (SRH), including unintended pregnancies, and AYA also face unique challenges in uptake and continuation rates of LA contraceptives. LA contraceptives, including injectable and implantable methods, are increasingly used and frequently co-delivered within HIV programs. Additionally, AYA are highly interested in co-delivery of ARVs and contraceptives. Yet, co-delivery raises potential pharmacological, behavioral, and implementation issues needing further investigation. We propose foundational pharmacokinetic (PK) and qualitative studies leading up to a longitudinal study among AYA receiving LA injectable ARVs in Kenya, Botswana, and Alabama. Our central premise is that use of LA ARVs will foster long-term thinking for SRH, including for pregnancy prevention, and that leveraging existing clinics already providing LA contraceptives will make offering LA ARVs highly feasible. Aim 1a will determine if combined injectable LA ARV use has any bidirectional drug-drug interactions with injectable or implantable contraceptives. Method: Parallel group PK evaluation with repeat plasma sampling among LA ARV users initiating injectable or implantable contraceptives vs. those not using any hormonal contraceptives (total 5 groups, n=21 per group). Aim 1b will qualitatively explore points of convergence and divergence, preferences and values, and health systems readiness around wider-scale co-delivery of LA ARVs/contraceptives. Method: 2-4 focus group discussions with patients, providers, and policymakers. Aim 2 will evaluate the impact of co-delivery of LA ARVs and contraceptives among AYA via a longitudinal study on: (a) effectiveness outcomes of HIV (uptake and adherence/persistence) and contraception (uptake and continuation rates), and (b) implementation outcomes of acceptability and feasibility. Method: 6-monthly longitudinal follow-up of AYA receiving LA ARVs in multiple clinics in Botswana and Alabama over 2 years (estimated total n=1050).

NIH Research Projects · FY 2025 · 2021-04

ABSTRACT Human immunodeficiency virus 1 (HIV-1) is a major global pathogen, with 1.7 million new infections in 2019. The HIV-1 envelope glycoprotein (Env) is the sole virus-surface antigen. Env, a trimer of gp120/gp41 dimer subunits, is densely glycosylated by N-glycans. For the gp120 subunit, N-glycans contribute over 50% of its molecular mass and form a protective surface, the so-called “glycan shield”, that interacts with cellular receptors and the immune system and protects the virus from neutralizing antibodies (nAbs). Recently, the concept of an “evolving glycan shield” has been proposed, based on the sequence analysis of immune-escape variants that have lost some potential N-glycosylation sites (NGS) and gained others. Despite significant strides in understanding the biology of HIV-1 Env, the complexity and function of N-glycans at a molecular level are poorly understood. Based on our bioinformatics and experimental studies, we recently proposed that the Env glycan shield may be defined, and analyzed, as distinct structural glycan-microdomains. Using naturally occurring Env variants of a clade B virus, we analyzed the glycans surrounding the highly conserved N262 glycan, in the so- called high-mannose patch (HMP) near the apex of the Env trimer. These HMP glycans, comprised of different combinations of 4 to 6 NGS in the proximity of N262, appear to form a glycan working unit. Based on our published results and preliminary data, we hypothesize that Env outer domain, gp120 glycan shield, has structural components that can be defined as distinct structural glycan-microdomains. We propose to test our hypothesis in different clades of HIV-1 and their immune-escape variants by assessing structural and functional impacts of a range of N-glycan combinations that comprise Env glycan-microdomains. Specifically, we propose to determine how different NGS combinations of the N262-anchored HMP-microdomain across variants and clades influence Env functions (Aim 1), to probe the structural arrangement of different N262-glycan cluster NGS combinations in the context of functional recombinant Env trimers (Aim 2), and to assess the functional impacts of other potential glycan-clusters of the HIV-1 Env glycan shield (Aim 3). To accomplish these goals, we will use a combination of functional, bioanalytical, structural bioinformatics, and molecular dynamics simulation approaches. The results of these studies will test our proposed hypothesis on the functional impact of the structural components of the glycan shield, glycan-microdomains, and will determine how the varied compositions of these microdomains affect the function of the glycan shield. These data will advance our understanding of the molecular biology of HIV-1 and the factors that impact viral transmission and persistence. Ultimately, these studies will generate new information about Env vulnerabilities that can be exploited to develop future therapeutic approaches.

NIH Research Projects · FY 2025 · 2021-04

Project Summary/Abstract Approximately 50% of the 3.4 million persons with epilepsy (PWE) in the U.S. have cognitive problems with memory deficits reported most commonly. Memory deficits negatively contribute to the personal and professional outcomes of PWE including quality of life. While cognitive rehabilitation programs have been utilized to help improve memory function in epilepsy, their efficacy remains unclear. Recently, memory improvements with formal exercise training have been shown in randomized controlled trials in healthy individuals, and in our pilot study in PWE who participated in supervised combined endurance and resistance training. Despite evidence of memory and other cognitive benefits derived from physical exercise, clinical trials of exercise as a rehabilitative strategy for memory improvement in PWE have not been conducted to date. Further, we lack understanding of the brain changes that occur with exercise and their role in improving memory function as well as the longitudinal effects of exercise on memory and brain changes. Improving our understanding and ameliorating the negative impact of epilepsy, seizures, and its treatment are recognized by the epilepsy, medical and scientific community as an important focus of research. Thus, exercise training is a promising rehabilitative strategy for mitigating the memory deficits experienced in epilepsy. The primary objectives of this study are to determine in adults with idiopathic generalized epilepsy (IGE) the efficacy of a 6-week supervised and structured exercise program combining endurance and resistance training for memory rehabilitation, investigate a putative mechanisms of action for exercise-related memory benefits, and determine if the memory benefits and brain changes are retained 6 weeks after completing the exercise intervention. Based on our pilot exercise data in adults with epilepsy, we hypothesize that exercise will significantly improve verbal memory function in the exercise group compared to the no-intervention control group. We also hypothesize that the verbal memory improvements are mediated by the changes in resting state functional connectivity of the hippocampus, a brain region that plays a vital role in memory function. We propose a mediation model in which exercise-induced changes in the hippocampus functional connectivity is mediating the beneficial effect of exercise on memory function in epilepsy, and we will utilize the causal-steps approach in which 4 conditions of statistical significance must be met to determine if mediation is present. Using a randomized delayed intervention study design, we will conduct a clinical trial of the 6-week supervised exercise program combining endurance and resistance training in adults with IGE in order to test our hypotheses. Completion of the proposed study will significantly impact the field in that it will lay the groundwork for the use of exercise for memory rehabilitation in epilepsy, in which option for combatting memory impairment are limited.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY/ABSTRACT This K99/R00 application describes the background and experience of the applicant, Rachel Wells, PhD, RN, and her plan to acquire the knowledge and training necessary to become a leading independent clinical investigator in developing and testing early palliative care interventions that optimize the outcomes of underserved individuals living with advanced heart failure (HF). This training and proposed research are of importance because: 1) substantial unmet palliative care needs of persons living with advanced HF; 2) few investigators are examining the mechanisms underlying palliative care intervention components to optimize intervention effect; and 3) the development of efficacious and scalable palliative care interventions in HF is a key research priority. The overall K99 goals are to gain advanced research training in developing behavioral interventions, conducting randomized controlled trials using the Multiphase Optimization Strategy (MOST), and advanced research ethics to refine and pilot test components of a lay navigator-led early palliative care intervention for underserved persons with advanced HF in the Southern U.S. The goal of the UPHOLDS intervention (Utilizing Palliative Care for Heart Failure Optimized using Lay Navigators to Decrease Suffering), will be to improve quality of life (QOL) by activating advanced HF patients through trained lay navigators coaching. The specific training objectives during the K99 phase are to: (1) develop and tailor interventions that are: 1a) manualized with rigorous fidelity procedures for enhanced replication and scalability; 1b) tailored to underserved populations; and 1c) led by lay navigators; (2) develop skills to design and lead RCTs using the MOST framework; and (3) continue training in research ethics. A comprehensive training plan has been developed in concert with mentorship team of senior research experts and includes intensive face-to-face mentorship, formal coursework/workshops, clinical trial observerships, and conference attendance. The K99 research aim will be met using interviews with a) underserved persons with advanced HF, b) lay healthcare navigators, and c) palliative care and HF clinicians, to modify and refine the content, format and delivery of HF palliative care intervention components. The overall goal during the R00 phase is to conduct an optimization pilot using a 23 factorial design to assess acceptability, feasibility, and potential efficacy of the newly refined UPHOLDS intervention. The research specific aims during the R00 phase are to: 1) determine the feasibility and acceptability of using a highly innovative MOST factorial design approach to enroll and retain 40 advanced HF patients for 24 weeks and 2) explore the preliminary efficacy of individual intervention components and component interactions on patient outcomes (QOL, primary outcome) at 12- and 24- weeks after baseline. The results will directly support an R01 application to conduct a fully powered MOST factorial trial to test the effects of individual palliative care intervention components on advanced HF patient outcomes.

NIH Research Projects · FY 2024 · 2021-04

Project Summary  Calcium  oxalate  stone  disease  occurs  in  nearly  10%  of  the  U.S.  population  and  contributes  significantly  to  health care costs and negatively impacts quality of life. The amount of oxalate excreted in urine is a known risk  factor for calcium oxalate stone disease. Approximately 50% of urinary oxalate is derived from the diet and the  remaining  from  endogenous  synthesis.  The  metabolism  of  ascorbic  acid  (AA),  an  important  antioxidant,  is  a  source  of  urinary  oxalate  derived  from  endogenous  synthesis.  However,  the  contribution  of  this  source  to  urinary oxalate excretion is not well defined. Prior experiments have been hampered by 1) a lack of control of  dietary  oxalate  to  urinary  oxalate  excretion  and  2)  the  confounding  generation  of  oxalate  from  AA  in  non-­ acidified  urine  samples.  The  turnover  of  AA each  day  is  approximately 80  mg and  could  feasibly  result  in  the  formation  of  up  to  40  mg  of  oxalate  per  day.  We  have  measured  the  contribution  of  AA  turnover  to  urinary  oxalate  excretion  with  carbon-­13  labelled  AA  oral  dosing  in  a  small  number  of  normal  adults  and  have  confirmed  that  AA  contributes  40  -­50  %  of  the  endogenous  oxalate  excreted  in  urine.  These  preliminary  findings  suggest AA  turnover  is  a  major  source  of  urinary oxalate derived  from  endogenous  synthesis.  In this  proposal  we  will assess  the  conversion of  AA  to oxalate  in non-­stone  forming  adults  and  CaOx  stone  forming  adults  using  the  stable  isotope  of  AA,  carbon-­13  AA.  We  will  further  examine  the  effects  of  obesity,  which  is  known  to  be  associated  with  systemic  oxidative  stress  and  a  decreased  plasma  AA  concentration,  on  this  conversion as well as increased oxalate excretion. Subjects in nutritional studies will ingest known amounts of  food-­derived AA  in  diets also  controlled  in  their  contents  of oxalate,  calcium and  other  nutrients. Experiments  will be conducted in cultured cells and mouse models to systematically examine the relationships between AA  and  oxalate,  the  role  of  pro-­oxidants  in  this  process,  the  role  of  mitochondria  and  AA  transport  into  this  organelle,  and  to  determine  whether  antioxidants  can  blunt  oxalate  formation  from  AA.  If  these  studies  are  successful they  will  open  new  avenues  of  research  for  decreasing urinary  oxalate excretion and  kidney  stone  formation.

NIH Research Projects · FY 2026 · 2021-04

PROJECT ABSTRACT Epileptic encephalopathies (EEs) are severe brain disorders of early infantile and childhood age onset characterized by epileptic seizures, abnormal electroencephalogram (EEG), severe cognitive and behavioral impairments that might lead to early death. It is estimated that ~2.9 million Americans live with epilepsy and the mortality rate in people with epilepsy is ~2-3 times higher than the general population. Several genetic mutations associate with EEs including mutations in the X-linked intellectual disability gene CASK that are found in patients with Ohtahara syndrome (OS) and West Syndrome (WS). Constitutive CASK deletion in mammals is incompatible with life and the prognosis of CASK hemizygous male patients remains extremely grim. The precise function of CASK and the potential mechanisms by which CASK mutation produces EE remains obscure. Because the constitutive CASK-/- knockout mice exhibited neonatal lethality, we recently generated a novel mouse model of EE by deleting CASK specifically from the neurons (CASKNKO). We found that CASKNKO mice display severe growth retardation, recurrent tonic spasms, EEG anomalies, and myoclonus beginning postnatal day 17 that leads to death by postnatal day 25. Multiple studies have shown that CASK protein is localized at the mitochondrial membranes. Recently, CASK gene expression was found to be regulated in an NAD+/Sirtuin1 dependent manner in mouse neurons. Moreover, we found that mammalian CASK interacts and co-localizes with mitochondrial proteins, and significantly modulates mitochondrial function and number. Based on the evidences from literature and our findings we hypothesize that CASK plays a role in brain mitochondrial function and metabolism, and is critical for optimum neuronal excitability in vivo. To test this hypothesis, we will examine the brain mitochondrial, metabolic, and electrophysiological functional changes as well as synaptic excitatory/inhibitory balance in the CASKNKO mice. We will further identify the specific domain/s of CASK that interacts with mitochondrial proteins, and determine if SIRT1-dependent mitochondrial biogenesis pathway is dysregulated in the brain of CASKNKO mice. Experiments will be performed before and after the onset of myoclonus to distinguish between a potential cause and consequence relation with the disease. We will also test if pharmacological activation of NAD+/SIRT1 pathway can stimulate mitochondrial biogenesis in the brain and CASK expression in glial cells to rescue EE phenotype in the CASKNKO mice. Success in the proposed project will uncover how loss of neuronal CASK alters mitochondrial and synaptic functions to produce EE. The long-term goal of our project is to use the novel CASKNKO EE mouse model to identify potential disease biomarkers and test therapeutic strategies for clinical intervention.

NIH Research Projects · FY 2024 · 2021-04

Abstract Long-range (>10 μm) transport of electrons along networks of G. sulfurreducens protein filaments, known as microbial nanowires, has been invoked to explain a wide range of globally important redox phenomena. The remarkable electronic conduction capability of those nanowires has sparked a great deal of interest in the medical application space, such as for building biocompatible materials and biosensor. For more than a decade, G. sulfurreducens nanowires were thought to be bacterial type IV pili, supported by many indirect genetic and biochemical observations. Recently we showed that these conductive nanowires are not made of type IV pilins. Instead, these structures are a polymerized multi-heme c-type cytochrome, OmcS, which have never been characterized before. The OmcS filament model is consistent with the known roles of OmcS in Geobacter respiration, but our knowledge of cytochrome appendages is still very limited. This study aims at addressing fundamental scientific questions about cytochrome filaments in respiring prokaryotes as well as applying our discoveries into the general medical field. Specifically, I will: A) identify and characterize novel cytochrome filaments in bacterial and archaeal strains, through bioinformatics algorithms followed by microscopic validation. B) Then I will study the conduction mechanism of these filaments by high resolution cryogenic electron microscopy (cryo-EM) and conductivity measurement. C) Finally, based on these new insights into cytochrome filaments, I will create a novel design for a self-assembled conductive nanowire. These nanowires may be derived directly from a novel cytochrome filament or may contain a peptide/protein based self-assembled scaffold core with soluble cytochromes appended to the outer surface. The results will advance our understanding of cytochrome nanowires, as well as generating self-assembling nanowire scaffolds that may be used in many future biomedical applications.

NIH Research Projects · FY 2024 · 2021-04

PROJECT SUMMARY/ABSTRACT This research proposal has been developed to equip the candidate, Dr. Daniel Tyrrell, with knowledge, experience, and skills to successfully transition into an independent faculty position. The proposed research will examine mechanisms of why older adults are at increased risk of vascular disease. Vascular pathology in the brain is present in up to 80% of people with Alzheimer’s disease (AD) and vascular dementia is the second leading dementia behind AD. Dr. Tyrrell will have guidance from his mentors, Dr. Daniel Goldstein and Dr. Michael Wang, and assistance from members of the Michigan Alzheimer’s Disease Research Center along with experts in diverse fields who will contribute to Dr. Tyrrell’s training. Dr. Tyrrell has discovered that cerebral blood vessels in aged mice have evidence of mitochondrial dysfunction leading to increased STING-IL-6 inflammation and blood-brain barrier breakdown. The mechanisms of vascular contributions to cognitive impairment and dementias (VCID) in aging remain unclear. Therefore, the objective of this study is to determine how cerebrovascular mitochondrial dysfunction impacts cognitive function in aging. Hence, Aim 1 will determine how age-related cerebrovascular mitophagy and mitochondrial dysfunction impacts the blood-brain barrier and cognitive function during natural aging and in a model of vascular dementia induced by cerebral hypoperfusion. Aim 2 will examine how cerebrovascular inflammation impacts the BBB and cognitive function in a model of vascular dementia with aging. Dr. Tyrrell will employ novel mitophagy reporter mice and mice with enhanced mitochondrial function in addition to using cell-type specific conditional knockout mouse models and pharmacologic approaches. Dr. Tyrrell will acquire both technical and scientific neurobiology expertise in order to study blood-brain barrier integrity and cognitive function in aging to distinguish himself from his mentors and allow him to develop a cutting-edge independent career in examining VCID.

NIH Research Projects · FY 2025 · 2021-04

Project Summary: The overall goal of this project is to provide applicant J. Nicole Bentley, MD appropriate resources and mentorship to develop skills allowing a path to research independence in the field of neuromodulation for cognitive disorders. This proposal focuses on the role of dorsolateral prefrontal cortex (DLPFC) in altered cognition in Parkinson’s disease (PD) patients with simultaneous intracranial cortical and subcortical recordings during awake deep brain stimulation (DBS) surgery. Cognitive impairment is a common non-motor symptom of Parkinson’s disease (PD), affecting one-quarter of newly diagnosed patients, and more than 80% of patients over the course of the disease. Despite its prevalence, very few treatment options exist. Unfortunately, mainstay treatments for motor symptoms of PD such as dopaminergic medications and high- frequency deep brain stimulation (DBS) do not improve and in some instances can even worsen cognitive symptoms. Recent evidence in PD patients suggests that reduced prefrontal theta frequency oscillatory activity is associated with impaired cognition, specifically evident in the domain of inhibitory control. Given its dense connections to basal ganglia and pre-motor areas, neuromodulation of the DLPFC-basal ganglia network could enhance elements of cognitive dysfunction in PD. Our novel preliminary data shows DLPFC and subcortical power modulation and phase-amplitude coupling during an intraoperative Simon task measuring inhibitory control, and that bursting theta-frequency DBS modulates DLPFC theta power. Here we propose studies to (1) simultaneously record DLPFC and globus pallidus interna (GPi) local field potentials (LFPs) in awake PD patients during the Simon task, and (2) contrast continuous and intermittent theta stimulation from the DBS electrode at the GPi to determine the effect on behavioral and electrophysiologic measures. Given the increasing prevalence of cognitive impairment and the dearth of available treatments, there is an urgent need to foster the development of clinician-neuroscientists with this interest. As a neurosurgeon with specialization in neuromodulation, Dr. Bentley is an ideal candidate to serve in this role. She has assembled a multi-disciplinary and multi-institutional team of expert mentors who will provide guidance and support towards the scientific and career development aims of this proposal. She and her mentors have designed a career development plan consisting of cognitive neuroscience, computational neuroscience, and signal processing techniques. These training aims will be combined with the high volume of a tertiary movement disorders functional neurosurgery practice at the University of Alabama, Birmingham. With the institution’s robust resources in research computing, biostatistics, engineering, and neurophysiology, this proposal will ensure that Dr. Bentley is well-prepared for progressing to research independence by the completion of this award.

NIH Research Projects · FY 2026 · 2021-03

PROJECT SUMMARY / ABSTRACT Patients diagnosed with cancer between the ages of 15-39y have not seen the same improvement in survival when compared with patients diagnosed as children (<15y). These patients have received special designation by the NCI as adolescents and young adults (AYA), accompanied by a mandate to address these outcome disparities. Acute lymphoblastic leukemia (ALL) epitomizes this phenomenon, where survival rates in AYA continue to lag behind children (5y overall survival: 1-14y: 86%; 15-21: 56%; 22-29: 42%; 30-39: 35%). Across pediatric-inspired and adult-style therapy regimens, ALL therapy includes a prolonged (~2y) maintenance phase of chemotherapy requiring daily oral 6-mercaptopurine (6MP). A large investigation recently confirmed that inadequate systemic exposure to 6MP is associated with an increased risk of relapse; to measure systemic 6MP exposure, this comprehensive approach assessed the levels of a 6MP metabolite (thioguanine nucleotide [TG]) incorporated into DNA (DNA-TG). Several AYA-specific factors likely contribute to systemic 6MP exposure and have not been examined systematically. These factors are likely related to both the AYA patient (6MP adherence) and healthcare (disease management, reflected by the provider-prescribed 6MP dose intensity [DI]), as well as the barriers and facilitators within these domains. We hypothesize that systemic exposure to 6MP will correlate with both 6MP adherence (in the patient domain) and disease management, as represented by provider- prescribed 6MP DI (in the healthcare domain). We will examine these associations between DNA-TG levels and both 6MP adherence and 6MP DI, adjusting for pharmacogenetics. We will then identify barriers/ facilitators to both 6MP adherence and disease management, using a mixed methods convergent parallel study design, both quantitatively determining the predictors of adherence and 6MP DI, and qualitatively describing the perspectives of both the patients and healthcare providers. We will conduct this study using a prospective cohort of AYA patients in ALL maintenance therapy across a national 14-site consortium of multiple types of facilities and oncology practices. For 6 months per patient, 6MP adherence will be monitored electronically and monthly DNA- TG levels will be followed (peripheral blood). Patient questionnaires will capture self-reported barriers and facilitators of adherence as well as sociodemographics; institution-level questionnaires will capture healthcare factors. Findings from this study have the potential to identify both patient-related and healthcare-related targets amenable to interventions with the potential to mitigate AYA outcome disparities in AYA patients with ALL. Furthermore, this study establishes the infrastructure to continue to follow this cohort for relapse, and conduct intervention(s) informed by this proposed trial within a novel AYA-focused consortium.

NIH Research Projects · FY 2026 · 2021-03

Project Summary The proposed R01 aims to augment data from the FORCE-TJR national Registry with radiographic readings, geospatial information and Medicare claims data to quantify timeliness of total knee replacement (TKR), and investigate the social determinants (SD) affecting timing of TKR. We propose 3 specific aims. Aim 1 applies validated appropriateness criteria to a representative sample of the FORCE-TJR registry patients to classify them as either premature or appropriate but with 5 different levels of functional disability. Aim 2 appends this sample with geospatial information (SD) about their environment and determine the effect of the SD factors on TKR utilization. Aim 3 determine the effect of the SD factors on improvement after surgery. We examine the effect of SD factors related to (1) the medical care environment (i.e., access to and practice patterns of medical care) and (2) the built environment (i.e., all of the physical parts of where we live and work (e.g., homes, buildings, streets, open spaces, and infrastructure)). Our project will be the first to examine timing of TKR at a national level, and will provide new knowledge regarding the association of the medical care and built environments with timeliness of TKR utilization. Our findings will also inform clinical and public policy interventions aimed at targeting TKR to those who would benefit most from the procedure, and add to evidence that would support the proposition that improvements to the environment could lead to an overall reduction in disability pertaining to knee osteoarthritis.

NIH Research Projects · FY 2025 · 2021-03

Project Summary/Abstract Retinitis pigmentosa (RP) is a heterogeneous group of disorders that cause retinal degeneration. To date, mutations in more than 58 genes have been linked to classic retinitis pigmentosa, and collectively all forms of inherited retinal dystrophies involves at least 300 genes. My studies are focused on retinitis pigmentosa-59 (RP59), a non-syndromic autosomal recessive form of RP caused by mutations in the dehydrodolichyl diphosphate synthase (DHDDS) gene, localized to exon 3 on chromosome 1p36.11. The most prevalent mutation is K42E, while T206A and R98W also have been reported. We generated a murine Dhdds K42E knock- in mouse model using CRISPR/Cas9 gene editing technology. Preliminary data shows significant reduction of ERG b-wave amplitudes and we have reported marked GFAP up-regulation, without retinal degeneration or decreased protein N-glycosylation. This suggests that defective DHDDS-dependent retinal degeneration may be more complicated than simply a loss-of-function mechanism due to altered protein glycosylation. To better understand the basis for selective retinal pathology associated with RP59 mutations, we plan to characterize the DhddsK42E/K42E knock-in mouse anatomically and physiologically. Additionally, we will create and analyze two new models of DHDDS-related inherited retinal disease, Dhdds T206A/T206A and DhddsT206A/K42E. Studying the retina in these mouse models will give us insight into the pathophysiology of this inherited disorder, and may lead to novel treatment options for this disorder.

NIH Research Projects · FY 2025 · 2021-03

PROJECT SUMMARY/ABSTRACT Dr. Sayer’s career goal is to become an independent translational obesity scientist at the University of Alabama at Birmingham (UAB) with a research program focused on the development of pragmatic and adaptive weight loss interventions that can be rapidly implemented in clinical and community settings. The proposed K01 training plan and research project will provide the knowledge, skills, and abilities to launch his independent research career in pursuit of his long-term goals. Dr. Sayer’s training plan includes a combination of formal didactic training available through the UAB Center for Outcomes and Effectiveness Research and Education (directed by Dr. Kenneth Saag, Co-Mentor) as well as rigorous hands-on and personalized training through the NIH National Rehabilitation Research Resource to Enhance Clinical Trials (directed by Dr. Marcas Bamman, Co-Mentor), UAB Nutrition Obesity Research Center (directed by Dr. James Hill, Primary Mentor), and UAB Comprehensive Center for Health Aging (directed by Dr. Cynthia Brow, Co-Mentor). The proposed K01 research is a pilot Sequential Multiple Assignment Randomized Trial (SMART) using a high protein diet and resistance training to promote the retention of fat free mass during weight loss in men with obesity aged 50 and older. The primary objectives of the research are to provide critical preliminary data on 1) the feasibility of conducting a full-scale SMART, 2) estimates of intervention effects and their variances, and 3) moderators or predictors of intervention responsiveness that may serve as tailoring variables for a full-scale SMART. Dr. Sayer will leverage data and training resulting from the proposed K01 to successfully compete for R01-level research funding to develop and rigorously evaluate a full-scale SMART designed to construct an optimal adaptive treatment strategy using a HP diet and RT for preventing and/or treating sarcopenic obesity.

NIH Research Projects · FY 2025 · 2021-02

PROJECT SUMMARY/ABSTRACT Chronic myelogenous leukemia (CML) results from hematopoietic stem cell (HSC) transformation by the BCR- ABL tyrosine kinase. Tyrosine kinase inhibitors (TKI) are effective in inducing remission and prolonging survival in CML patients, but fail to eliminate primitive leukemia stem cells (LSC) that can regenerate disease. Most patients need ongoing TKI treatment to maintain remission, and remain at risk of toxicity, financial hardship and non-adherence. The long-term goal of our research is to improve understanding of mechanisms of LSC resistance to treatment, to support development of effective and safe strategies for LSC targeting, and enhance possibilities of treatment-free remissions in CML patients. Mitochondrial metabolism plays a critical regulatory role in normal HSC function. CML LSC demonstrate increased mitochondrial oxidative phosphorylation (OXPHOS) compared to low OXPHOS in normal HSC. However, mitochondria also play important roles in metabolic processes besides OXPHOS, including fatty acid, glutamine and glucose oxidation, and generation of biosynthetic intermediates. The rationale for our studies is that specific mitochondrial metabolic alterations that contribute to altered LSC growth and TKI resistance are not known. Our preliminary studies show initial inhibition of OXPHOS in CML LSC after TKI treatment, but subsequent restoration of OXPHOS, and increased fatty acid oxidation (FAO), with continued treatment. A SIRT1, P53 and MYC regulatory network plays an important role in LSC propagation. We show that SIRT1 and its target PGC-1α play an important role in increased OXPHOS in CML LSC. PPARa, a PGC-1α-coactivated transcription factor and a key regulator of FAO, shows increased expression in CML LSC after TKI treatment, and contributes to increased OXPHOS, proliferation and survival. We will explore the hypothesis that increased FAO following BCR-ABL kinase inhibition, together with maintenance of high levels of OXPHOS, glycolysis and glutaminolysis, contributes to TKI resistance in CML LSC, and that metabolic regulatory mechanisms represent potential targets for elimination of TKI-treated CML LSC. In Specific Aim 1 we will use a combination of gene expression, extracellular flux, metabolite profiling and in vitro and in vivo metabolic labeling to study effects of TKI treatment on mitochondrial metabolism in CML LSC, examine the role of SIRT1, PGC1a and PPARa in metabolic alterations, and study interactions of MYC and p53 regulatory networks with mitochondrial metabolism. In Specific Aim 2 we will investigate the role of increased OXPHOS and FAO in promoting TKI resistance in CML LSC. Bone marrow microenvironment niches play a critical role in maintaining quiescent, TKI-resistant LSC populations. However, the role of the microenvironment in metabolic regulation of LSC growth is not known, and will be evaluated here . These studies are significant since they are expected to identify mechanisms of metabolic regulation underlying TKI resistance in CML LSC, establish connections between metabolism and other regulatory mechanisms in CML LSC, and identify new targets for therapy. The concepts developed here will have broad implications for other malignancies.

NIH Research Projects · FY 2025 · 2021-02

PROJECT SUMMARY Hepatocellular carcinoma (HCC) accounts for nearly 31,000 deaths annually in the United States alone. Factors that epigenetically silence an HCC tumor suppressor gene have the potential to promote tumorigenesis and thus may provide novel drug targets for HCC therapies. Using a genome-scale shRNA screen, we identified BCL6 as a putative and novel HCC driver gene. BCL6 is a transcriptional repressor with no previously documented role in HCC development and progression. We found that BCL6 was sufficient to transform cultured immortalized hepatocytes and promote HCC tumor growth in mouse subcutaneous xenografts. These oncogenic effects of BCL6 were dependent upon the ability of BCL6 to cause transcriptional repression because a transcriptional repression activity defective mutant of BCL6 failed to transform hepatocytes and promote tumor growth in mice. Based on our preliminary results, in this application we will establish the role of BCL6 as a driver of HCC, determine the mechanism by which BCL6 promotes tumor growth, and evaluate novel BCL6 targeting small-molecule inhibitors for HCC therapy. In Aim 1, we will establish the role of BCL6 in initiation and progression of hepatic tumorigenesis using a series of complementary mouse models that recapitulate characteristic features of HCC, including a mouse model of liver fibrosis (cirrhosis). This model recapitulates cirrhosis which is a cardinal feature of HCC. Additionally, based on our preliminary results that BCL6 inhibition in HCC cells increases their probability of getting cleared by T-cells, we will also use a mouse model with humanized immune system to study the impact of BCL6 in HCC progression in the context of a functional human immune system. In Aim 2, based upon the results of our RNA-seq and further follow up analysis, we will determine if epigenetic silencing of tumor suppressor HHIP and the zinc transporter ZIP14 by BCL6 is necessary for it to drive hepatic tumor growth. In Aim 3, we will evaluate our novel BCL6 inhibitors in pre-clinical mouse models of HCC, including HCC patient-derived xenograft (PDX) models for HCC therapeutics. In preliminary experiments, we have developed a novel small-molecule inhibitor designed to target the BTB domain of BCL6 and thereby disrupt its interaction with its co-repressors SMRT, NCOR and BCOR. We found that this novel BCL6 inhibitor L2-12019 inhibited the HCC cell growth in culture and in a human HCC xenograft-based mouse model. In Aim 3 studies, we will rationally design and evaluate new L2-12019 analogs to improve drug-like properties. The lead analog (with improved potency, selectivity, stability and solubility) will be tested with L2-12019 in pre-clinical mouse models of HCC and its efficacy will be compared with existing BCL6 inhibitors. Collectively, the results of the experiments proposed in this application will elucidate a novel non-genetic druggable pathway that promotes HCC tumor growth and progression and evaluate a new approach for treating HCC.

NIH Research Projects · FY 2024 · 2021-02

Project Summary/Abstract Candidate: My long term goal is to become an independent investigator running an interdisciplinary and collaborative research team to understand the mechanisms underlying the effects of medications and drugs of abuse on intra/interconnected brain circuits . Specifically, my interests surround understanding how cholinergic interneurons (ChAT) regulate the activity of nicotinic acetylcholine receptors (nAChRs) in the local circuitry of the nucleus accumbens (NAc), thereby underlying sex differences in dopamine signaling associated with reward and motivated behavior. I have a strong background in electrophysiology and pharmacology, and propose to be trained in optogenetics, behavioral pharmacology, and in vivo fiber photometry to round out my training. This will provide me with the skills to produce high impact publications and successful R01 submissions. I received my PhD in April of 2017 and this is currently my third year of postdoctoral research training. Training: In addition to Dr. Calipari, I have an advisory committee of experts in academic research who will provide the necessary training and guidance to accomplish this proposal. Dr. Barnett, is the Vice-chair of and a Professor in the Department of Pharmacology and Dr. McCabe is the Director of the Office of Postdoctoral Affairs. Outside of the committee, we have identified, courses, seminars, and meetings to provide further technical training, presentation experience, responsible conduct in research, and the necessary skills (negotiations, tenure, laboratory management, etc.) to transition to independence. Research: Substance use disorder (SUD) is a chronic, recurring brain disease characterized by significant dysfunction in reward-seeking behavior. A large body of work has focused on understanding the neural mechanisms in the brain’s reward circuitry, yet these studies have overwhelmingly focused on male subjects. Epidemiological evidence shows that women represent a particularly susceptible population to SUD, yet the mechanisms in the brain that underlie sex differences in reward and motivation are largely unknown. The neural control of reward is dependent on dopamine release in the NAc that is subject to heavy modulation by ChAT signaling through nAChRs; a process that is essential to encoding information about environmental reward predictive cues. My preliminary data show fundamental sex differences in the regulation of dopamine release via nAChRs, yet to date, it is not known how this process occurs, what factors influence this effect, and how this relates to motivated behavior. The overall goal of this proposal is to define sex-specific circuit-based mechanisms governing sex differences in reward processing. By using voltammetry techniques along with pharmacology, behavioral analysis, and in vivo dopamine recording, I anticipate being able to expand our understanding of the sex differences in ChAT regulation of nAChR modulation of dopamine release underlying reward learning. The successful completion of this proposed project has the potential to inform the development of better and more effective pharmacotherapies to counter neurological disease states in women.

NIH Research Projects · FY 2026 · 2021-02

Protocol Title: A dental implant registry of treatment outcomes of implant therapy by practitioners in the National Dental Practice Based Research Network Summary There is a substantial body of literature to support that biological and prosthetic complications occur which may interfere with the health of the peri-implant tissues, the function and esthetics of the implant restoration. Peri- implant diseases are classified into peri-implant mucositis, inflammation restricted to the peri-implant mucosa, and peri-implantitis, characterized by peri-implant bone loss. The limitations of the current body of literature of biologic and prosthetic complications are based on many small studies and in large part conducted in an academic and specialty setting. We propose to create an implant registry within the National Dental Practice Based Research Network (NDPBRN) that will record the setting and implant therapy, the implants used, the prosthetic therapy provided and the rate of complications. The registry will create an opportunity for subsequent, additional targeted studies on specific complications available from the registry data and will lead to diagnosis driven therapy strategies. The target enrollment is a total of 2000 implants with prosthesis across the whole network. The one year UG3 Phase will be used to create a strategy to be able to meet the recruitment and enrollment objectives and to develop the protocol for data collection to facilitate the data gathering of the practitioners for the subjects they enroll. The data collection model we propose will be designed to be validated, concise, and easy to use for practitioners. We will develop a web based decision tree that will guide the practitioner through the data collection. The UH3 phase will recruit practitioners with representation of all 6 regions of the network that will enroll subjects with 2000 implants. Data collection will be detailed and comprehensive and will include surgical, prosthetic, and biologic aspects of implant therapy and radiographs for bone level assessments for a period of 3 years. The data will present clinically meaningful information about the prevalence of the various implant therapies, the incidence of prosthetic and biologic implant complications, risk factors for implant complications and evidence-based implant therapy strategies in every dental practices. We expect that the results from this study will significantly impact the clinical practice of implant dentistry and the quality of care provided for the patients.