University Of Massachusetts Amherst

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY Caspases are cysteine proteases that control apoptotic cell death. Caspases are activated to kill cancerous cells, but inhibiting caspases can prevent deleterious cell death in diseases like heart attack and stroke. Thus, there has been significant interest in caspases as drug targets. This interest heightened further with the finding that caspase-6 plays a central role in neurodegeneration. Unfortunately, no caspase-directed therapies are on the market, primarily because research has focused on the active site, the most conserved region of the family. It is clear that each caspase is regulated in a unique manner. A comprehensive understanding of which is essential to achieve caspase-specific inhibition. Thus, our long-term project goal has been to define and exploit unique regulatory features for each apoptotic caspase. Our pursuit of that goal has been successful. Due to our understanding of the unique features of each apop- totic caspase, we developed an allosteric caspase-6 inhibitor that is more potent than any reported (33 nM) and is also by far the most selective, preferring caspase-6 500-fold or more over all other caspases. This selectivity was possible because the allosteric site we targeted is unique to caspase-6, locking it into a helical conformation not attainable by any other caspase. It is gratifying that our intense and systematic study of caspase regulation - cleavage state, conformational change, zinc binding and phosphorylation - culminated in a structural understanding that enabled us to meet our goal of caspase-selective allosteric inhibition. Thus, we propose research that will further extend our understanding of the regulation of the apoptotic caspases in new key areas - substrate selectivity by caspase isoforms, core unfolding and aggregation of caspase-9 by phosphorylation, and interactions between caspase core and prodomains to achieve substrate selection. In addition, we are currently moving this caspase-6 inhibitor forward toward clinical use. While we are thrilled at the success of developing an inhibitor that can block the function of just one of the twelve human caspases, the substrate-selective inhibitors proposed here promise to be even more invaluable. For example, preventing caspase-6 cleavage of Tau would have a major impact on preventing the formation of neurofibrillary tangles in Alzheimer’s Disease. On the other hand, cleavage of DJ-1 (PARK7) by caspase- 6 is important for preventing Parkinson’s Disease. Thus, an ideal caspase-6 inhibitor for treatment of Alz- heimer’s disease would block Tau cleavage, but would still cleave apoptotic substrates and DJ-1, without promoting cancer or Parkinson’s disease. The proposed studies identifying exosites for Tau and DJ-1 will enable us to develop substrate-selective inhibitors (nanobodies) that can block cleavage of Tau, but not of DJ-1. Together this work plan increases both our fundamental understanding of the biology and regulation of caspases and enables development of a novel highly tuned class of caspase-directed therapies.

NIH Research Projects · FY 2026 · 2023-05

Project Summary HIV and food insecurity pose severe and interrelated problems in Latin America and the Caribbean, including in the Dominican Republic (DR), where HIV ranks as one of the top 5 causes of death and our prior studies have found that nearly 70% of people with HIV (PWH) have moderate or severe food insecurity. Despite the established, detrimental role of food insecurity on poor HIV treatment outcomes, evidence on sustainable interventions that address the cycle of food insecurity and poor HIV health is scarce. To address this gap, we developed and piloted ProMeSA (through an R34 grant), an integrated urban gardens and peer nutritional counseling intervention, and found it feasible, acceptable, and with preliminary efficacy at 6 and 12 months of improving food security and HIV virologic suppression. The purpose of this five-year study is to conduct a fully- powered cluster randomized controlled trial (RCT) of ProMeSA to assess intervention efficacy evaluated over a longer period (18 months) as well as mediators and barriers and facilitators to intervention uptake, implementation, and sustainability. The specific aims are: (1) Determine the efficacy of an integrated urban gardens and peer nutritional counseling intervention on the primary outcome of HIV viral suppression [undetectable HIV viral load (VL)] and secondary outcomes of ART adherence and HIV care retention care among people with food insecurity across diverse regions in the DR; (2) Examine the intervention effects on intermediate outcomes posited to mediate the impact of ProMeSA on ART adherence, care retention, and viral suppression; (3) Evaluate process-related factors associated with intervention uptake and implementation (facilitators, barriers, fidelity, and replication costs) to inform future scale-up. The trial will include 20 HIV clinics randomized to intervention or usual care control (n=25 per clinic; 500 total study participants). VL and other key outcomes will be assessed at baseline, and 6-, 12- and 18-months. Following our intervention causal framework and pilot findings, we hypothesize that ProMeSA will improve food security and diet quality and reduce stigma and competing needs, which in turn will improve HIV clinical outcomes. The predominant causal paths identified will inform tailoring ProMeSA to enhance impact in future dissemination and implementation. In addition, we will collect extensive quantitative and qualitative data on intervention implementation and participant experiences with the intervention across diverse settings and participants to inform scale-up. The study involves a partnership among researchers from the University of Massachusetts Amherst, University of California, San Francisco, RAND, and the Universidad Autonóma de Santo Domingo as well as the Dominican Ministries of Agriculture and Public Health, the Dominican National HIV/AIDS Council, and the United Nations World Food Program. To our knowledge, this will be the first full-scale trial to integrate nutritional counseling with food-generating activities among PWH with food insecurity, in support of national and international goals to achieve viral suppression and reduce the disease- and economic burden of HIV.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY/ABSTRACT Infertility and pregnancy loss are common and associated with significant morbidity. Ambient air pollution may affect reproductive health through alterations in the inflammatory response and increased production of reactive oxygen species, leading to cellular damage, epithelial dysfunction and platelet activation. Prior research has identified associations between air pollution exposure and population-level fertility; however, interpretation is limited by an incomplete understanding of underlying pathophysiologic processes and the reproductive windows most susceptible to air pollution exposure. This exposure misclassification risks underestimating the impacts of air pollution on women’s reproductive health and studies refining exposure windows timed to underlying biologic mechanisms remains a pressing need. This exploratory/developmental research grant proposal will leverage high-quality secondary data from a completed preconception time-to- pregnancy cohort, the Effects of Aspirin in Gestation and Reproduction (EAGeR) trial (n=1,228), to evaluate the impact of acute exposure to ambient air pollution during susceptible windows of the menstrual cycle and in early pregnancy on women’s reproductive health and ability to achieve a live birth. We will utilize Community Multiscale Air Quality (CMAQ) models to estimate participants’ residential exposure to ambient air pollution, including the criteria pollutants, constituents of particulate matter, and polycyclic aromatic hydrocarbons. The fine spatial and temporal resolution of the air quality data will allow for assessment of acute exposure during biologically-informed windows of the menstrual cycle (preovulatory follicle development, ovulation, and implantation) and early pregnancy. Aim 1 will evaluate the impact of acute exposure to ambient air pollution and PAHs around ovulation and implantation with alterations in reproductive hormones and fecundability. Aim 2 will evaluate the impact of acute exposure to ambient air pollution and PAHs from implantation through the first trimester on risk of pregnancy loss. For both aims, we will analyze mid-cycle biospecimens in a subset of participants (n=288) to assess metabolites of exposure to polycyclic aromatic hydrocarbons and oxidative stress mechanisms, including markers of oxidative DNA damage (8-hydroxy-2’-deoxyguanosine) and lipid peroxidation (F2-isoprostanes). We will additionally leverage the unique design of the EAGeR trial to identify whether randomization to the anti-inflammatory and anti-platelet actions of low-dose aspirin mitigates the impact of air pollution exposure on women’s reproductive health. These findings will provide the foundation for future work evaluating the mechanisms through which acute exposure to ambient air pollution may impact the ability of couples and individuals to achieve a live birth, both informing policy development and identifying future intervention points to improve reproductive health.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Despite tremendous advances in treatment of HIV/AIDS and the decrease of HIV incidence, the overall infected population continues to grow. Progress on prevention of HIV transmission remains far too slow. It is estimated that 20% of new HIV infections are due to transmission from unaware infected individuals. Hence, early detection of HIV is particularly important for lowering transmission rates. To this end, extending testing accessibility beyond clinical settings through self-tests is highly desirable. HIV self-testing is a process in which an individual who wants to know his/her HIV status collects a specimen, performs the test, and interprets the result in private. Current HIV self-testing technologies include rapid protein tests and nucleic acid tests. The suboptimal sensitivity of current protein tests can only support antibody detection and will miss a significant portion of acute infections. Although nucleic acid tests can reach lower detection limit through amplification technologies such as PCR, they will miss the information that antigens can provide. Most current research efforts on HIV self-testing are focused on nucleic acid tests. However, there is no evidence showing that HIV RNA appears ahead of antigen. The major technological challenge for antigen detection is that proteins cannot be amplified like nucleic acids, leading to the widely held belief that antigen tests are relatively insensitive and therefore have a limited clinical utility. We previously demonstrated a click chemistry amplified nanopore (CAN) assay method for ultrasensitive antigen quantification. This assay achieved 0.5 pg/ml detection limit for HIV-1 p24 antigen and demonstrated reliable detection in clinical samples from patients missed by nucleic acid and/or ELISA assays. Quantitative p24 results from this method also indicated correlation between p24 and viral load, suggesting potential use for monitoring antiretroviral therapy adherence to minimize treatment failure. Based on the CAN assay, this project aims to develop an ultrasensitive quantitative HIV-1 p24 antigen self-test to improve early detection of acute infections and monitoring treatment efficacy. The test will be based on a streamlined automatic device including a cost- effective microfluidic chip for sample preparation and a nanopore reader for laypersons to test themselves using fingerprick blood. The R61 phase will develop a self-testing device for quantification of HIV-1 p24 antigen at ≤ 2 pg/ml in whole blood to support detection of acute infection and treatment failure. The R33 phase will perform a clinical evaluation at the Prisma Health Immunology Center to assess the performance, usability, and acceptability of the self-testing device. Through innovations in click chemistry amplified nanopore detection and microfluidic sample preparation, we anticipate the test would be able to quantify HIV-1 p24 antigen at as low as 0.5 pg/ml directly from 100µl or less finger prick blood and establish correlation between p24 and viral load levels. If successfully developed and validated, this CAN self-testing device should enable routine HIV self-testing as simple as a blood sugar test at home to support global HIV diagnostic and therapeutic efforts.

NIH Research Projects · FY 2025 · 2023-01

ABSTRACT Despite tremendous advances in treatment of HIV/AIDS and the decrease of HIV incidence, the overall infected population continues to grow. Progress on prevention of HIV transmission remains far too slow. It is estimated that 20% of new HIV infections are due to transmission from unaware infected individuals. Hence, early detection of HIV is particularly important for lowering transmission rates. To this end, extending testing accessibility beyond clinical settings through self-tests is highly desirable. HIV self-testing is a process in which an individual who wants to know his/her HIV status collects a specimen, performs the test, and interprets the result in private. Current HIV self-testing technologies include rapid protein tests and nucleic acid tests. The suboptimal sensitivity of current protein tests can only support antibody detection and will miss a significant portion of acute infections. Although nucleic acid tests can reach lower detection limit through amplification technologies such as PCR, they will miss the information that antigens can provide. Most current research efforts on HIV self-testing are focused on nucleic acid tests. However, there is no evidence showing that HIV RNA appears ahead of antigen. The major technological challenge for antigen detection is that proteins cannot be amplified like nucleic acids, leading to the widely held belief that antigen tests are relatively insensitive and therefore have a limited clinical utility. We previously demonstrated a click chemistry amplified nanopore (CAN) assay method for ultrasensitive antigen quantification. This assay achieved 0.5 pg/ml detection limit for HIV-1 p24 antigen and demonstrated reliable detection in clinical samples from patients missed by nucleic acid and/or ELISA assays. Quantitative p24 results from this method also indicated correlation between p24 and viral load, suggesting potential use for monitoring antiretroviral therapy adherence to minimize treatment failure. Based on the CAN assay, this project aims to develop an ultrasensitive quantitative HIV-1 p24 antigen self-test to improve early detection of acute infections and monitoring treatment efficacy. The test will be based on a streamlined automatic device including a cost- effective microfluidic chip for sample preparation and a nanopore reader for laypersons to test themselves using fingerprick blood. The R61 phase will develop a self-testing device for quantification of HIV-1 p24 antigen at ≤ 2 pg/ml in whole blood to support detection of acute infection and treatment failure. The R33 phase will perform a clinical evaluation at the Prisma Health Immunology Center to assess the performance, usability, and acceptability of the self-testing device. Through innovations in click chemistry amplified nanopore detection and microfluidic sample preparation, we anticipate the test would be able to quantify HIV-1 p24 antigen at as low as 0.5 pg/ml directly from 100µl or less finger prick blood and establish correlation between p24 and viral load levels. If successfully developed and validated, this CAN self-testing device should enable routine HIV self-testing as simple as a blood sugar test at home to support global HIV diagnostic and therapeutic efforts.

NIH Research Projects · FY 2026 · 2023-01

Slightly over 1 in 5 children in the United States are growing up in a household that speaks a language other than English. For these children, developing bilingual skills will allow them to communicate both with family members who have limited English skills and with English-speaking teachers and peers. However, for children with autism spectrum disorder (ASD), there is limited research about how to facilitate their bilingual development. Professionals and parents have often assumed that learning two languages would be too challenging, especially for children with minimal spoken language skills. While several studies over the past decade have demonstrated that bilingual exposure is not detrimental for language development in children with ASD, these studies have focused mostly on the acquisition of English rather than examining factors that promote development of both languages. Furthermore, bilingual studies have generally excluded children with minimal/low verbal skills. The proposed sequential mixed-methods project places a new emphasis on variability within the bilingual experiences of children with ASD and their families. It seeks to identify factors associated with children’s skills in both languages, as well as their social-cognitive skills such as perspective- taking and cognitive flexibility. The quantitative phase will include 60 Spanish-speaking families with children with ASD (ages 4-6), including children who are minimally verbal. The first aim is to characterize sources of variation in the types of bilingual language environments experienced by children with ASD (e.g., different languages from caregivers vs. therapists; different languages from different caregivers; code-switching by bilingual caregivers; changes in language choices over time). The second aim is to examine predictors of children’s dual-language communication and social-cognitive skills, including the role of quantity and contexts of Spanish exposure. In a qualitative phase conducted with a subset of 30 families, the third aim is to interpret the quantitative findings in reference to family perspectives, priorities, experiences, and challenges with language choices and support services. The long-term goal of this work is to collaborate with families and service providers to develop interventions that support bilingual families and promote the linguistic and social- cognitive development of children across the autism spectrum. This career development proposal includes an expert team of mentors from Psychology, Communication Disorders, and Public Health. The training goals focus on conducting research with minimally verbal children with ASD, mixed methods, and using community- based participatory research to develop and evaluate interventions. The research aims and training goals of this career development award reflect current priorities in the strategic plan of the NIDCD for Voice, Speech, and Language, including Improving Diagnosis, Treatment, and Prevention in Understudied Populations (Priority Area 3) and Improving Outcomes for Human Communication through community-based research (Priority Area 4).

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Cell to cell communication is critical for function in all multicellular organisms. A key factor for intercellular communication is regulation by Ca2+ concentration. Ca2+/calmodulin dependent protein kinase II (CaMKII) is a Ca2+ sensitive enzyme that is encoded by four genes in mammals: α, β, γ, and δ. There is an incredible amount of diversity generated from the four vertebrate CaMKII genes. Alternative splicing produces up to 386 transcripts, which leads to the production of 386 proteins that are then differentially post-translationally modified, and mix to form hetero-oligomeric complexes, ultimately culminating in thousands of chemically distinct CaMKII proteoforms. We are specifically interested in the crucial roles CaMKII plays in long-term memory formation (neurons: α, β), fertilization (oocytes: γ), and cardiac physiology (cardiomyocytes: δ). Intriguingly, these cells all communicate using Ca2+ oscillations but on vastly different timescales (minutes to milliseconds). How does one enzyme accommodate this multifunctionality? We hypothesize that selective splicing and modification creates a unique set of CaMKII variants expressed in specific cell types, thereby leading to differential functional outputs. Fully elucidating these complex biological roles requires a deeper understanding of CaMKII variation at the sequence and protein level, structural and conformational ramifications of these variations, and how these variables affect CaMKII interactions within the cell. In this proposal, we seek to expand our understanding of CaMKII function inside cells using a combinatorial approach of sequencing, biochemistry, structural biology, and cellular assays. Completion of the proposed work will allow us to uncover the molecular basis for the many roles of CaMKII in neurons, cardiomyocytes, and oocytes – with far-reaching implications on therapeutic intervention for neurologic disease, cardiac dysfunction, and infertility.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract The long-term objective of this program of research is to increase access to hearing health care for underserved older adults. The aims of this proposal focus on the hearing and communication needs of an older bilingual Latinx population. Aim 1 will focus on speech understanding in noisy backgrounds. We will investigate the impacts of age, hearing, cognitive processing, and bilingual language history on speech-in-noise performance. This aim includes lab-based and virtual test protocols that will measure speech understanding, nonspeech auditory processing, and nonverbal cognitive (i.e., executive function) processing skills. The results of these studies will improve our theoretical understanding of speech perception abilities for all older adults, while specifically increasing the knowledge base related to Spanish/English bilingual older adults. The investigations undertaken in Aim 2 will focus on assessing and defining the challenges associated with age, hearing loss, and bilingualism in every day communication. A mixed methods approach will be undertaken in Aim 2. One study will rely on ecological momentary assessment (EMA)—brief surveys sent via mobile device periodically throughout the day—to quantify the demand for use of both languages in a Latinx community of older adults within the broader context of a majority English-speaking environment. In addition, interviews with the EMA participants will add context to the survey questions related to communication and effort experienced in daily encounters. Finally, focus groups will explore issues related to aging and hearing loss, and their impact on social engagement for older bilingual Latinx adults. Importantly, we will seek to determine what the hearing and communication priorities are for this growing and underserved population of older adults. The knowledge generated regarding speech understanding and communication needs and priorities will be used in future grant applications to develop community-integrated service models that will better serve older adults.

NIH Research Projects · FY 2025 · 2022-09

Summary There is a worldwide shortage of kidneys for transplantation due mainly to the fact that there is no reliable means to determine the viability of kidneys available for transplant. There are unmet clinical needs to reduce the >8,000 deaths that occur each year from failures of finding viable kidney matches for transplant, to reduce the nearly 4- year time on donor waitlists, and to reduce the number of failed transplants. The current process for screening deceased donor kidneys uses two methods: 1) pathological scores based on anatomical features from a biopsy (tubules, glomeruli, etc.) and 2) the Kidney Donor Profile Index (KDPI) derived from the donor’s medical history (hypertension, diabetes, weight, etc.). However, clinical research indicates that those current methods have limited discriminative power. Optical coherence tomography (OCT) is an imaging technology that can obtain high-resolution, non-invasive, cross-sectional images of biological tissues in situ and in real time. We have demonstrated that OCT can provide non-invasive, real-time, histopathological information of the kidney that is impossible to obtain using any other known procedure. We have demonstrated that OCT imaging of human kidney histopathology both prior to and following their transplant can be used to predict post-transplant renal function. Furthermore, these preliminary trials have demonstrated that OCT imaging of human donor kidneys with a hand-held unit in the operating room is safe and that the entire kidney can be evaluated within a relatively short period of time. From a recently finished clinical study with 169 human transplant kidneys, we found that in the expanded criteria donor (ECD) kidneys (or marginal kidneys), increased tubular lumen diameter was able to predict delayed graft function (DGF) prior to implantation. In this proposal, we will develop a novel OCT device with intelligent scanning and deep learning to evaluate donor kidney viability before transplant. By scanning the whole kidney surface, our device intends to eliminate the uncertainty created by the biopsy/KDPI paradigm. The OCT imaging studies will be correlated with post-transplant renal function in order to establish imaging algorithms and guidelines for OCT imaging of kidneys prior to their transplant. Our central hypothesis is that more comprehensive morphological parameters as measured by OCT can be used to determine post-transplantation renal function. The specific aims of this proposal are: 1) Develop robot-assisted automatic 3D scanning OCT imaging device to image human kidneys prior to their transplantation. 2) Develop deep-learning-based image processing algorithms to quantitatively assess the OCT parameters as indicators of the functional status of kidneys, using histology scoring system as the gold standard. 3) Derive the diagnostic criteria for assessing transplant kidney function and perform prospective clinical studies to assess the accuracy of predicting post- transplant function using OCT.

NIH Research Projects · FY 2025 · 2022-09

Pregnancy complications are emerging as important predictors of future cardiovascular health with the demands of pregnancy acting as a “stress test” that reveals a pre-disposition to future cardiovascular disease (CVD) in the mother. Pregnancy may also reveal a predisposition to future depressive disorders, a hypothesis that is consistent with the co-occurrence of, and bi-directional relationship between, depression and CVD, particularly in women. Therefore, the long-term objective of this research is to understand how physical and mental health during pregnancy can help predict CVD events and mental health disorders in later adulthood. The specific objective of the proposed research is to examine the association of pregnancy complications and prenatal mental health with subsequent cardiometabolic profile and mental health in middle adulthood among Hispanics of Puerto Rican heritage living in the continental US. The overarching hypothesis of this proposal is that pregnancy complications will be associated with poorer cardiometabolic health and mental health status in middle adulthood in Puerto Ricans, a Hispanic subgroup at high risk of CVD. Currently, the majority of evidence regarding these associations derives from retrospective cohort studies that linked national birth and death registry databases in Europe. No studies have evaluated these associations in Puerto Ricans despite the fact that this group has the highest prevalence of diabetes, obesity, and major cardiometabolic risk factors among Hispanics and a population growth rate three times higher than the general US population. The proposed study provides a unique opportunity to leverage our previously-collected prospective pregnancy data from Proyecto Buena Salud (PBS, R01 DK064902, PI: Chasan-Taber). PBS was a prospective study of Puerto Rican women conducted from 2006-2011 that provided novel evidence that pregnancy complications and prenatal depression were highly prevalent in Puerto Ricans and were significantly associated with adverse maternal and birth outcomes. It has now been an average of 13.4 years since the participants’ PBS pregnancy. Our feasibility study projects that 87% (n=1,096) of PBS participants will participate in our proposed follow-up study. For the proposed study, bilingual/bicultural staff will conduct in- person visits to collect biomarkers of insulin resistance, inflammation, lipids, adiposity, blood pressure, and chronic physiological stress (hair cortisol concentrations). Interviewer-administered questionnaires, validated in Hispanics, will measure depression, psychosocial stress, and anxiety. We will use actigraphy to evaluate the novel hypothesis that poor sleep and low physical activity, highly prevalent among Puerto Ricans, may mediate the above relationships. This research is significant in prospectively assessing whether pregnancy complications offer a meaningful opportunity for early CVD prevention efforts. This research also has translational significance in informing culturally sensitive prenatal interventions for early life prevention of future chronic disorders in an understudied and particularly vulnerable population.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Several human proteins are known to form amyloid fibrils, and these fibrils are associated with devastating diseases, including Alzheimer's, Parkinson's, type II diabetes, and dialysis- related amyloidosis (DRA). The protein β-2-microglobulin (β2m) forms amyloid fibrils in the joints of patients undergoing hemodialysis, leading to DRA. While protein amyloid formation has been extensively studied, molecular-level information about the early stages of amyloid formation is only beginning to be revealed for a few proteins. This information, though, is critical for the rational development of therapeutics against amyloid diseases, particularly for DRA that has no treatment. Our group has been developing and applying new mass spectrometry (MS) methods to reveal structural and oligomeric changes that β2m undergoes before forming amyloid fibrils. Such structural information is very challenging to obtain using traditional biophysical methods, and the structural insight from our new methods has led to exciting discoveries about the first steps of protein amyloid formation. We are beginning to utilize this insight to find inhibitors of DRA. The research in this MIRA project seeks to (i) develop new methods that will deepen our mechanistic insight into the first steps of β2m amyloid formation and (ii) leverage this new mechanistic information to discover robust inhibitors of β2m amyloid formation. These goals will be accomplished by creating new MS methods that rely on covalent labeling, ion mobility, and computational modeling. These new approaches will allow us to probe the energy landscape of the structural switch that initiates β2m amyloid formation, providing unprecedented quantitative insight into the factors that cause this normally stable protein to become amyloidogenic. We will also explore new methods to characterize the on- and off-pathway isomeric oligomers that are present during the early stages of β2m amyloid formation, allowing us to reveal the specific protein-protein interactions that lead to amyloids. Altogether, the structural insights that we obtain will then be utilized to discover new inhibitors of β2m amyloid formation. Specific outcomes of this research will be new biophysical tools to study protein amyloid formation and the identification of inhibitors that could lead to therapeutics against DRA. The universality of the techniques developed in this work will also make them applicable to other amyloid systems and diseases.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY There is clinical and scientific interest in developing on-demand local anesthesia, in which local anesthetics can only be released by an external trigger when and where needed after the first administration. In the past few years, injectable local anesthetic formulations have been developed that can generate on-demand local anesthesia triggered by light and ultrasound. However, their clinical application is hindered by shortcomings, such as basal drug release when the trigger is not applied, a small number of repetitive trigger events for one injection (usually 4 to 6 times), the trigger event only occurs within a few hours after the injection, potential local tissue toxicity caused by the external triggers, and expensive preparation procedures. In this research, in order to overcome these shortcomings, we aim to explore cold triggered local anesthesia. The formulation design consists of three components: tetrodotoxin (TTX), cold therapy, and thermo-responsive polymersomes. We hypothesize that TTX is an extremely potent local anesthetic, a small amount can successfully produce local anesthesia, thereby improving the efficacy of each trigger and increasing the number of trigger events; cold treatment is a common use of hypothermia in medical therapy, which has the characteristics of low cost, easy operation, and non-invasiveness; thermo-responsive polymersomes can efficiently encapsulate TTX at body temperature, but release TTX due to the deconstruction of the polymersomes under cold treatment. In addition, the micron-sized polymersomes can stay at the injection site for a long time, acting as a drug reservoir to achieve long-term effects. Specifically, a large library of polymersomes with various structures (size and porosity) and properties (structurally stability and shell stiffness) will be fabricated. Such a library will be a collection of candidates. Iterative screening will be conducted to identify the polymersomes with the desired in vitro cold triggerable TTX release, enhanced local retention in tissues, and excellent biocompatibility (cytotoxicity, inflammation, myotoxicity, organ toxicity). Then, the rat footpad anesthesia model and rat paw incision wound infiltration model will be used to evaluate the in vivo efficacy and safety of the selected polymersomes in cold triggered local anesthesia. The expected outcome of this project is a convenient and applicable on-demand local anesthetic formulation. Such a system will enable patients to adjust the degree of local analgesia according to their changing needs and conditions, and will minimize the administration of systemic analgesic medications such as opioids.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract This proposal describes the development of tools that will be applied to robustly investigate the role of circadian rhythms in tissue engineering for the first time by: synchronizing rhythms in vitro; reinstating absent rhythms locally in regenerating sites in vivo; and aligning therapeutic delivery to distinct circadian phases. Our lab focuses on timing as a tool in tissue engineering, whereby we use therapeutic signals to co-ordinate various phases of repair over the time scale of days. But, 24 hr circadian rhythms – which play critical roles in biological systems (e.g., tissue formation, cell signaling, cell activity) – are ignored in tissue engineering studies and product development. For example, in current tissue engineering cell cultures individual cells have a circadian rhythm, but they rapidly fall out of synch with other cells in the culture, eliminating rhythmic processes that would occur in the natural system. Indeed, patient populations with disrupted circadian rhythms (e.g., diabetes, obesity) also have poor tissue regeneration and they are the target of many tissue engineering strategies. Yet, nobody has considered this lack of rhythmicity in developing tissue engineering strategies. From the therapeutic timing perspective, there is strong evidence in many diseases that the efficacy of systemically delivered drugs is strongly affected by the time of day. In local tissue regenerating sites, it is highly probable that there are also optimum circadian times for therapeutic delivery, which needs to be investigated. In the next five years we will begin to fill these gaps in tissue engineering research laying the groundwork for an unexplored approach that can be harnessed to optimize tissue regeneration. First, this work will impart the currently absent circadian rhythms into in vitro tissue engineering studies of skin, bone, and vascularization. This will be accomplished using a robotics device that interfaces with standard tissue culture setups to provide synchronizing signals (e.g., glucocorticoids, temperature) at precise timepoints. Interestingly, glucocorticoid rhythms are altered in patient populations with poor regeneration (e.g., obesity, diabetes) but synthetic glucocorticoids given systemically can reinstate circadian rhythms. In a first-of-its-kind approach, we will test the ability to drive local circadian rhythms at a regenerating site via glucocorticoid signaling. To do this we will develop circadian drug delivery systems based on ultrasound- or light-triggered release that are capable of precisely timed daily local delivery of glucocorticoids. The automated tools and circadian drug delivery system will additionally be modified and used to demonstrate the effect of growth factor alignment with circadian phases on tissue repair. This proposal will rigorously investigate the interplay between circadian rhythms and tissue engineering, provide the tools to research and test these rhythms, and demonstrate how these rhythms can be used to enhance regeneration. With this foundation, our labs’ vision is to lead exploration of this new facet of tissue engineering, undertaking future work that manipulates these rhythms for regeneration of multiple tissues, more deeply explores the mechanisms, and translates these new tools and findings to the clinic.

NIH Research Projects · FY 2025 · 2022-08

Project Summary/Abstract In the U.S., heart failure (HF) affects over 6 million people and is one of the most common causes of hospitalization. New technologies are needed to enable in-home monitoring to guide and treatment changes for patients at risk of developing Acute Decompensated HF (ADHF). Early detection of decompensated HF often relies on monitoring weight gain, but weight alone does not accurately gauge the fluid accumulation that predicts worsening of HF. A device that can measure vital signs (including heart rates and respiratory rates), intrathoracic fluid status (using thoracic bioimpedance), and heart rhythm, may allow for more accurate identification of the early stages of acute decompensation in chronic HF patients. The multidisciplinary team from the University of Massachusetts (UMass) Amherst, UMass Medical School, and the University of Connecticut proposes to develop a novel device for in-home monitoring of HF patients who are at risk of decompensation. The central hypothesis is that an innovative bioimpedance and electrocardiogram monitor with re-usable, non-wetted, and flexible bioimpedance electrodes embedded in a wearable vest, used in conjunction with a smartphone and cloud server, will continuously collect, transmit, and monitor key physiologic data. Devices running decision-support algorithms will analyze these data to identify patients with an emergent HF decompensation that may be mitigated with prompt medical attention. A Bioimpedance and Electrocardiogram Device (BED) attached to the back of the vest collects 5 minutes of data once a day from the vest electrodes, calculates bioimpedance, and has fault- tolerant assessment circuits to enable reliable data collection. Collected data consisting of heart rhythm, some vital signs, intrathoracic fluid accumulation measurements, and information about the data reliability will be sent to a smartphone from the BED via Bluetooth-Low-Energy (BLE) wireless communication. This data, together with the patient's weight, and symptoms and signs noted in the application, such as shortness of breath or lower leg swelling, will be recorded by patients and be sent to a cloud server via 4/5G/WiFi mobile networks. These data will be used to develop a robust clinical decision support algorithm that accurately detects early ADHF. This project aims to: A1.1) develop a wearable vest with reusable carbon-black and polydimethylsiloxane (CB/PDMS) electrodes that capture 3-channel bioimpedance and electrocardiogram data; A1.2) develop a BED with fault- tolerant circuit; A1.3) develop a smartphone application, to include a machine learning algorithm that uses the collected physiological information for autonomous early ADHF detection; A2.1) establish a cloud infrastructure that allows for the collection of the aforementioned data from the in-home setting along with associated data reliability monitoring; and A2.2) evaluate the performance and usability of the system in a prospectively recruited cohort study of patients with known HF. The clinical study will target a diverse HF population that are at high risk for ADHF. A successful project will result in the design, testing, and clinical evaluation of a prototype telehealth monitoring system to collect real-time data about cardiac risk factors for people with HF.

NIH Research Projects · FY 2026 · 2022-07

Abstract: Inflammation is a defense mechanism triggered by innate immune system against any foreign invasion to restore homeostasis, but when it sustains for a prolonged period of several months to years, it transforms into a chronic condition resulting in several harmful diseases. The inflammasome is a hetero-multimeric protein complex known for activating inflammatory caspases followed by subsequent processing of cytokines, which makes it one of the key players during inflammation. Abnormal activation of inflammasomes can initiate undesirable inflammatory responses associated with the progression of chronic inflammatory diseases. Several studies have investigated nanomaterial interactions with immune cells to understand their role in various biological applications and tailor them to different needs. Indeed, several types of nanomaterials have been widely explored to target the immune cells at the disease site to modulate the immune responses. However, our recent studies and several recent reports suggest that many of these nanomaterials activate inflammasomes in immune cells non-specifically, potentially exacerbating the disease. But the comprehensive characterization of the nanomaterial-immune cell interactions that results in inflammasome activation and the unwanted innate immune response is poorly studied due to a lack of appropriate investigative tools. The overall vision of my research program is to design immunoengineering platforms bridging nanoscience and engineering design with manipulation of the immune system to address fundamental and translational questions in immunology. We focus on developing effective immunotherapy strategies by understanding the interactions between different immune system components, between various nanomaterials and immune cells. Specifically, we aim to address fundamental questions in inflammasome biology and how nanomaterial properties affect their interactions with innate immune cells and inflammasome activation. To accomplish this, we propose to engineer a library of multiparametric polymeric nanomaterial platform with various surface and core characteristics in a single system. This will allow us to test the effect of these nanomaterial properties on inflammasome activation, tweaking one property at a time to develop nanomaterial structure-property-function relationships. We have developed novel high-throughput imaging platform to enable monitoring of inflammasome activation in real time. We have also engineered novel imaging probes to monitor inflammasome activation in vivo in real time. Using these tools, over next five years, we aim to understand the interactions between nanomaterials (polymer-based) and immune cells (macrophages, monocytes, dendritic cells and neutrophils) in the context of inflammasome activation and uncover the mechanisms of this activation in vitro and in vivo. In summary, the information obtained from these studies could provide design criteria that guide the development of next-generation of nanomaterials to control, prevent or mitigate inflammasome signaling pathways and also provide a predictive framework for modulation of the inflammasome activation for potential applications in diagnostics and therapy.

NIH Research Projects · FY 2025 · 2022-07

In New England, where the predominant vector-borne disease (VBD) burden is due to ticks, tick control and suppression practices can barely keep up with regional endemicity conditions, despite the fact that New England has a long history with ticks. With just 4.5% of the U.S. population, New England accounts for 20% of confirmed Lyme disease cases in the U.S. This project proposes a New England Center of Excellence in Vector-Borne Diseases (NEWVEC) to make significant progress in combatting ticks and other arthropod disease vectors in the six New England states through applied research and technical evaluation of methods to prevent vector bites, suppress disease-causing vectors, and promote public adoption of vector-control measures. To sustain and amplify this public health imperative, NEWVEC will train students to enter the public health entomology workforce to address VBD, and will engage stakeholders through a community of practice that can help its research and training efforts remain relevant, effective, and impactful. The NEWVEC strategy has three components: Applied research on tick suppression, Training of public health entomologists with expertise in vector-borne diseases, and Community of Practice to enhance regional effectiveness of evidence-based methods. Applied research projects will follow four parallel streams: 1) Standardizing and optimizing personal protection and control products and applications for commercial and residential use; 2) Discovering and evaluating emerging technologies to suppress ticks and prevent tick biting; 3) Designing and testing habitat and host-targeted interventions for suppressing tick populations; and 4) Assessing human factors and public health approaches for tick control. Work in these areas will be conducted by researchers at six New England universities, supported by two core facilities: a Molecular Analysis lab for pathogen testing and taxonomic bar- coding at the University of Massachusetts Amherst (UMA), and a Tick rearing and experimental test bed at the University of Rhode Island. A multi-tiered training program in Public Health Entomology based at UMA will develop concentrations and certificate programs at both the undergraduate and graduate levels to train highly competent graduates for employment in the field, while providing a range of high-quality professional development training experiences for public health professional staff and technical employees of commercial applicator firms around New England. NEWVEC’s Community of Practice will promote extra-academic collaboration in its research, promoting two-way transmission of vital knowledge among the center’s researchers, its students and trainees, the public health stakeholder community, and others concerned with vector-borne disease prevention in New England. NEWVEC will concentrate on the most important New England vectors, most challenging barriers to VBD progress, and most promising and impactful vector control interventions and methods. By being technically credible, NEWVEC will be relied upon by the VBD practice community, thus influencing adoption of recommendations and promoting scalability to reach all areas in the New England region.

NIH Research Projects · FY 2026 · 2022-06

Several neurodegenerative diseases, such as Alzheimer’s disease (AD), are characterized by the spread and aggregation of the protein tau. Recently, we identified a cellular receptor, LRP1 (Low-density lipoprotein Receptor-related Protein 1), that regulates the tau spread pathway. Knockdown of LRP1 prevents tau spread in human iPS neurons and the mouse brain, suggesting that the tau-LRP1 interaction could be an important entry point for disease intervention. Unfortunately, a detailed understanding of the tau-LRP1 molecular complex is still lacking. Therefore, the main objective of this project is to define the tau-LRP1 structural interface and discern how post-translation modifications (PTMs) to tau’s structure influence tau uptake and spread. In preliminary work, we have developed protocols to purify and measure interactions between tau and LRP1. We have established cellular platforms to model tau propagation and have shown that this process can be influenced by tau PTMs. To fully develop this work, we propose three aims. In Aim 1, we will use TR-FRET to establish in vitro affinities between tau and LRP1 and mass spectrometry to map the protein-protein interface. In Aim 2, we will look at how tau phosphorylation can influence the tau-LRP1 complex and what effect this has on tau spread and aggregation. In Aim 3, we will focus on tau PTMs that alter lysine residues. We will assess if ubiquitination or acetylation can impact the tau-LRP1 interaction, if they influence tau aggregation, and if they promote or inhibit tau uptake and seeding in cells. The innovative experimental methods and comprehensive analyses outlined herein will provide important mechanistic insight and develop our understanding of pathogenic tau regulation in AD. This will be an essential first step forward for the development and evaluation of potential AD therapeutics.

NIH Research Projects · FY 2026 · 2022-03

Project Summary/Abstract Recent recognition of the prevalence of intrinsically disordered proteins (IDPs) in biology and human diseases has challenged the traditional paradigm that stable structure is required for protein function. Furthermore, many IDPs have been found to remain disordered even in specific complexes and functional assemblies. These discoveries have now dramatically expanded the meaning of “structure” in the protein structure-function paradigm, to include a continuum from disordered ensembles to well-defined conformations. Importantly, these disordered proteins and dynamic interactions are central components of the regulatory networks that dictate virtually all aspects of cell decision-making. They are associated with a growing number of human diseases including cancers, neurodegenerative diseases, diabetes and heart diseases. There is thus a crucial need to establish the molecular basis of how conformational disorder mediates protein function, so as to understand how these functional mechanisms may be perturbed in diseases, or rescued by drug molecules for therapeutics. The key challenge towards achieving these overarching goals is quantitative description of the disordered protein states in relevant biological and disease contexts. Experimental measurements of averaged structural properties alone are inadequate to define the disordered protein ensemble, and reliable molecular simulations have a crucial and transformative role to play. This project aims to continue to develop advanced molecular modeling and simulation methodologies that can provide accurate description of disordered protein states, expand the accessible time and length scales, and enhance our ability to embrace critical questions in molecular level biomedical research. Through strategically chosen experimental collaborations, this project will further tackle questions and problems centered around several systems of great biomedical significance: 1) To establish the sequence-structure-function-disease relationship of IDPs, we will determine how multisite phosphorylation and cancer-associated mutations modulate the structure, dynamics and interactions of the transactivation domain (TAD) of tumor suppressor p53; 2) To develop effective strategies for targeting disordered protein states, we will determine the molecular basis of how the anti-cancer drug EGCG inhibits p53-TAD through dynamic interactions and study the functional dynamics and inhibition of flaviviral proteases; 3) To understand dynamic protein-protein interactions in relevant contexts, we will determine the molecular basis of how molecular chaperone Hsp70 achieves selective promiscuity to help the cell cope with protein folding challenge and how a novel family of virulence protein named SPIN from S. aureus inhibits human myeloperoxidase for evading the host innate immune defense. Integrated computational and experimental approaches deployed throughout these studies will enable us to direct our computational method development efforts to critical areas for which advances are needed, while at the same time push and test our methods with tangible feedback. 1

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY/ABSTRACT Suicide is a leading cause of death worldwide, accounting for nearly 800,000 deaths each year.1,2 Suicide rates continue to increase in the U.S.3 Most research on suicide risk factors has identified static, dispositional factors.4 These data inform us about groups at actuarial risk, but do very little to predict with useful precision when people will have suicidal thoughts or act on suicide thoughts. Thus, there is an urgent need to identify proximal risk factors for suicide risk. Two of the leading factors distally associated with increased suicide risk are dysfunctional emotional responses5 and decision-making deficits.6 Suicidal individuals have poorer reinforcement learning,6,7 demonstrating impaired ability to learn and modify behavior in response to reward and punishment. Individuals at high risk for suicide (i.e., borderline personality disorder) also show difficulties learning from punishment and reward,8,9 and the PI’s data show that negative emotions uniquely impair learning in such high-risk populations relative to healthy and clinical controls.10 It is necessary to understand the interactive effects of emotion and reinforcement learning as factors elevating near-term risk for suicidal thoughts and behaviors. Therefore, the proposed study examines both emotional and cognitive processes associated with suicidal thoughts and behaviors among 170 patients with a range of suicidal risk recently treated in an emergency department for a suicidal crisis. Using innovative ecological momentary assessment methods (EMA), the proposed study will examine behavioral measures of reinforcement learning and both physiological and subjective measures of momentary emotions. Aim 1 identifies the main and interactive effects of emotional responses, reinforcement learning, and emotion-related decrements in learning on recent suicidal thoughts and behaviors among suicidal patients recently seen in the emergency department. Aim 2 examines how emotions and learning interact to predict momentary suicidal thoughts and behaviors over four weeks. Aim 3 tests emotion- related decrements in learning as prospective predictors of future suicide risk over a six-month follow-up. The research team (PI: Dixon-Gordon, Co-Is: Ammerman, Boudreaux, Rathlev; Consultants: Hackel; Collaborator: Laws) has access to world-class expertise, with extensive experience recruiting suicidal participants from medical settings, EMA, computational modeling, and experimental psychopathology. The most important challenge facing suicide prevention today is unraveling the complex emotional and cognitive proximal drivers that transform nascent suicide risk into action. Combining state-of-the-art behavioral measures and EMA, we will tease apart the complex longitudinal relations between dysfunctional emotional responses, reduced capacity to learn from reward and punishment, and fluctuations in suicidal thoughts and behaviors. Given the high societal costs of suicide, this work has important health significance. Findings will inform the development of treatments that target emotion dysfunction and learning in samples at-risk for suicide.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY: Overall More than 90% of older Americans would prefer to stay in their homes as long as possible as they age. However, the prevalence of chronic illness including Alzheimer's disease (AD) and Alzheimer's disease related dementias (ADRD) can make the goal of successful aging at home out of reach without substantial support. At- home health care technologies hold significant promise to provide many needed forms of support, but have not been specifically developed for older adults, AD/ADRD patients, caregivers, and their clinicians. Many existing health monitoring technologies are burdensome to use for older adults, are not sufficiently accurate, have specific algorithmic biases, and lack adequate usability, and the resulting deluge of data often fails to provide information that can usefully inform caregivers and support clinical decisions. Further, many current treatment and intervention regimes are limited in terms of their ability to be remotely delivered, managed and adapted to patient needs and caregiver abilities over time. The Massachusetts AI and Technology Center for Connected Care in Aging and Alzheimer's Disease (MAITC) is a multidisciplinary Center spanning five sites – the University of Massachusetts Amherst, Brigham and Women’s Hospital, Massachusetts General Hospital, Brandeis University and Northeastern University – that aims to foster interdisciplinary research on the development, validation and translation of emerging AI- enhanced technologies to enhance connections between older adults, caregivers, and clinicians in order to more effectively support healthy aging as well as the care of patients with AD/ADRD at home. To achieve this objective, MAITC will pursue the five aims. The first is to apply a rigorous process which involves eliciting stakeholder needs to drive the foci of pilot studies and identifying promising technologies suitable for incorporation into pilot research projects. The second is to form and support a multidisciplinary community of engineers, computer scientists, behavioral scientists, medical researchers, nurses and clinicians working to advance the goals of the MAITC. The third is to provide access to state-of-art validation facilities, diverse cohorts across rural and urban areas, and translation and commercialization services to enable rapid, robust and multi-faceted validation and translation of AI-enhanced technologies. The fourth is to administer a pilot project granting process and ensure that the pilot projects are responsive to stakeholder needs, use emerging AI and related technologies that have significant potential to improve interactions between older adults, lay caregivers, and clinicians. The fifth is to provide training to developers, clinicians, and other stakeholders and to disseminate findings to diverse research, development and practice communities to accelerate further development of promising technologies and promote the adoption of validated technologies beyond the MAITC. Through these coordinated efforts, we hope to improve the lives of older adults and their caregivers.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Human milk contains essential nutrients and bioactives that are transferred to the nursing infant in what was once considered in a linear manner. Though now there is considerable evidence that human milk directs early establishment of the microbiome through molecules such as oligosaccharides that modulate specific microbial populations. Microbial communities that colonize the gastrointestinal tract enter into a commensal relationship with their host potentially impacting physiology. Accordingly, nitrogen bound in urea is delivered at relatively high concentrations in breast milk and may be liberated for utilization by the host and commensals by microbial urease activity. We hypothesize that urea nitrogen salvaging (UNS) is a key syntrophic feature of host-microbial interactions early in life. This may be of particular importance to infants in this critical stage of development, or in host populations where dietary nitrogen is limiting. This hypothesis will be addressed experimentally by evaluating and characterizing the metabolic capacity for infant-associated commensals to utilize urea and transform it to a usable form by their host. Moreover, we propose to study infant microbiome- mediated UNS modeled in an in vitro model to identify community-level phenomena. By understanding the impact to community structure and function by urea metabolism, we will define hallmarks of a microbiome that performs UNS efficiently. This study investigates a poorly understood and hypothetical host-microbial interaction with implications to nitrogen homeostasis early in development. The inter-kingdom UNS pathway may be of critical importance to infants in general or in certain nutritional contexts. In addition, this study further defines what constitutes a protective infant microbiome based on aggregate community function. This would potentially inform diagnostics to assess UNS capacity as well as develop interventions to correct suboptimal UNS. As such, purposeful modulation of UNS would increase the repertoire of tools to direct microbiome function while personalizing for life stage, diet, and/or host phenotype.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Mammalian cells often use their actin cytoskeleton simultaneously for multiple functions, including cell motility, endocytosis, vesicular trafficking, and to establish and maintain cell polarity. Although they accomplish different tasks, functionally distinct actin networks share structural components and regulators that can include polymerization factors, polymer bundlers, and proteins that promote network turnover. This molecular multitasking by mammalian cells makes it difficult to determine how cells control subsets of actin functions. In contrast, budding and fission yeast have highly simplified actin networks that have been used to develop simple models of actin network control. The vast differences between yeast and human actin networks, howevxer, make it difficult to understand which principles of actin cytoskeletal regulation are shared. Chytrid fungi represent a natural bridge between the well-understood actin networks of yeast and the elaborate actin networks of human cells for three reasons. First, the evolutionary position of chytrids falls between yeast and human cells. Second, they have retained important actin regulators that have been lost by yeast, making their actin networks are intermediate in complexity. Third, and most importantly, chytrid fungi undergo a natural developmental transition from human-like crawling cells to a yeast-like cell type. We are therefore using chytrid fungi to study the evolution and specification of actin networks relevant to human health. We have identified a large number of chytrid actin cytoskeletal regulators similar to actin regulators found in human cells, as well as additional fungal-specific actin regulators. We have also recently developed methods to control the switch from the human-like to the yeast-like cells, as well as methods for molecular transformation and exogenous gene expression. We will use these approaches to determine the mechanisms that control animal-like and yeast-like actin networks in chytrids and the developmental transitions between them. This work will help us determine how actin regulatory systems give rise to the observed diversity of actin networks across developmental transitions and evolutionary time.

NIH Research Projects · FY 2026 · 2021-08

POJECT SUMMARY Traumatic brain injury (TBI) causes a number of deaths and permanent disability. Today 5.3 million people live with disability caused by TBI in U.S. Typically, TBI is initially assed by the Glasgow Coma Scale on hospital admission and routine neurologic examination. Computed tomography (CT) scanning and magnetic resonance imaging (MRI) are used for confirmatory testing. However, CT and MRI are not always available for every visitor for initial screening at emergency department; and there is no CT and MRI in the pre-hospital settings. Measurement of biomarkers in blood can provide an alternative, cost- effective and rapid route to screen TBI. In 2018, the U.S. Food and Drug Administration (FDA) authorized marketing of blood testing of two diagnosis biomarkers, ubiquitin c-terminal hydrolase-L1 (UCH-L1) and glial fibrillary acidic protein (GFAP), to evaluate TBI. Also, neuron-specific enolase (NSE) has been extensively investigated as a diagnosis blood biomarker for TBI. But there is no any commercial portable in vitro diagnostic tool to rapidly measure these biomarkers in blood in the emergency department and prehospital settings. Recently the PI developed a point-of-care (POC) prototype device that is able to rapidly measure TBI biomarkers in plasma and blood. To bring this prototype POC device to the clinical use, it will require at least three steps: (i) analytical validation; (ii) clinical validation; and (iii) FDA approval and 510(k) clearance. Analytical validation of the prototype device is the first step, which has not been done yet, but it is necessary to accomplish prior to clinical validation. The objective of project is to perform analytical validation of the paper-based lateral flow strip (PLFS) for detection of TBI diagnosis biomarkers in blood, in which surface-enhanced Raman scattering (SERS) is utilized for sensing signal transduction. Three types of PLFS will be used to measure three most recognized TBI diagnosis biomarkers including NSE, UCH-L1 and GFAP with assistance of a portable Raman detector, respectively. Each TBI biomarker can be detected individually with a small volume (around 35 µL) of blood sample within 36 minutes. With establishment of appropriate quality control and improvement procedures, the performance of PLFS will be validated analytically in terms of accuracy, precision, analytical sensitivity, reportable range, and selectivity. This application is in response to the NIH Program Announcement of PAR-18-550 (analytical validation) instead of PAR-18- 548 (clinical validation). If successful, such an inexpensive and rapid diagnostic POC tool will change practice in TBI diagnosis in the emergency department and pre-hospital settings. It will reduce unnecessary CT scans, increase the accuracy of TBI diagnosis, save costs, and enable earlier intervention aimed at mitigating both short and long term sequelae.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract Preterm infants are at increased risk of health complications due to their under-developed organ systems at birth, and human milk receipt is recognized as an important intervention to promote infant health. Recent identification of cells in human milk which have similar characteristics to stem cells, as well as animal models which show integration of these cells into nursling's organs, indicate an important function of milk cells which is currently not understood. In the NICU, infants often receive human milk which has been refrigerated or frozen, and then warmed and/or thawed prior to feeding. The impact of these typical handling practices on the cellular components of milk that may provide an important mechanism of biologic protection in infant health has not been investigated. This work will determine how the real-life storage and handling of human milk impacts infant health and development, and is the first to evaluate protective mechanisms. There are currently two major knowledge gaps in the fields of human milk and lactation and neonatal care surrounding bioactive cellular components. The first is a lack of knowledge of how real-life storage and handling practices impact the protective ability of milk's cellular components for infants. Without this knowledge, infants who may benefit from milk with the most bioactivity (such as preterm infants) may not receive the properties of their mother's milk which provide important protection. The second major opportunity is the lack of knowledge on the mechanism(s) of protection in milk stem-like cells. Additionally, as research techniques advance our ability to investigate milk components, there is a great need to look at milk components in a biological systems perspective. The specific aims of the research project are to 1) examine how hospital storage practices (refrigeration and freezing) impact the protective mechanisms of human milk cells through the use of a tissue culture model of intestinal health, and 2) determine if milk cells are integrated as functioning cells specific to vital organs impacted by preterm birth (brain, heart, lungs, intestine) using a cross-foster mouse model. Upon completion of the specific aims of the K23 research strategy and training plans, the candidate will have advanced theoretical knowledge and technical skills to conduct human milk research with the ability to apply a biological systems approach to understand the complexity of the many components of milk which likely impact the function of each. The proposal research aims are supported by research training, didactic coursework, scientific meetings, and specific plans for dissemination and future growth. Ultimately, the short- and long-term goals of this research are to 1) improve infant health by optimizing the delivery of the most bioactive human milk components and 2) determine the mechanism(s) by which human milk cells protect infants and promotes growth and development.

NIH Research Projects · FY 2024 · 2021-07

PROJECT SUMMARY Bladder reconstruction (BR) is essential to restore urinary function in patients with neurogenic bladder, congenital disorders, and as sequalae to the surgical treatment of bladder or pelvic malignancies. The current standard of care is to use the patient's own intestinal tissue (ileal segment; IS) as graft material during BR to create a neobladder or urinary diversion. While IS grafts are non-immunogenic and readily available, post- surgical persistence of intestinal cells in the graft impedes regeneration into bladder wall, resulting in stone formation, metabolic acidosis and risk of secondary cancers. Our objective is to develop new technology for intrasurgical tissue engineering of the IS by knockdown of cellular components using irreversible electroporation (IRE) and identify factors fundamental for regeneration of functional bladder wall. IRE is used in patients for tumor ablation by inducing cell death with ultrashort electric pulses. Our proposed strategy builds upon our preliminary data showing (i) IRE can knockdown intestinal cells in an IS graft, aiding repopulation with urothelium in a rat model of BR, (ii) feasibility of new pulse application strategies for the focal knockdown of mucosa, or decellularization while preserving vasculature and ECM in the IS, and (iii) phenotypic changes in IRE treated IS following urothelialization, that were not observed in sham controls under physiologic conditions of bladder filling and voiding. In specific aim 1, we will Define the impact of graft perfusion on bladder wall regeneration by performing vasculature sparing IRE of the IS. In specific aim 2, Elucidate the role of IS mucosa in post-BR complications by knockdown with IRE. In specific aim 3, Investigate the role of mechanotransduction in bladder function development in IRE treated IS. Intrasurgical creation of a perfused, histocompatible graft (Aim 1) and focal decellularization of the mucosa while sparing the underlying layers in the IS (Aim 2) are the first examples of in vivo knockdown tissue engineering using IRE. The study and application of mechanotransduction principles to augment urinary barrier function development in IS grafts (Aim 3) is previously undescribed. Knowledge gained from proposed research will yield a simple intrasurgical technique (mt-IRE or vs-IRE) that combines technology (IRE) and grafting technique (with IS) that are already in the clinic, enabling rapid translation for the immediate benefit of patients undergoing BR. Eventually, we anticipate our work to advance the concept of in vivo production of functionalized grafts using the patient as the source of biomaterial and the bioreactor, with application to reconstructive surgery involving other tubular organs such as the esophagus, trachea or large blood vessels.