Grants

NCI - National Cancer Institute

PROJECT SUMMARY Liver diseases affect nearly 4.5 million individuals in the United States. Clinicians utilize various imaging modalities (ultrasound, computed tomography, magnetic resonance imaging) to assess liver structure and presence of liver cancer. These modalities have a high degree of accuracy for identifying and characterizing liver cancer within the liver. However, the current imaging techniques lack the sensitivity for detecting metastatic tumors outside of the liver. Timely detections of these metastatic lesions have significant clinical implications relating to surgical management or liver transplantation. We have developed a liver-specific PET tracer which we hypothesize would provide a higher sensitivity for detecting tumors at a smaller size compared to the currently used imaging modalities. We anticipate that this tracer will also have non-oncologic applications such as assessment of liver function and biliary tract pathology. In this proposal, we aim to complete the preclinical testing and investigational new drug (IND)- enabling studies in preparation for first-in-human clinical trials.

NCI - National Cancer Institute

Summary MYC proteins (including c-MYC, N-MYC and L-MYC) are critical oncogenic drivers that regulate multiple cancer hallmarks. The evidence indicating that MYC is a critical oncogenic driver that regulates multiple cancer hallmarks is compelling. Recent pan-cancer analyses of over 9,000 human cancers of various types found amplification of MYC genes in ~28% of all cases. In addition, multiple mechanisms exist in tumor cells that frequently lead to the upregulation of MYC mRNA and protein expression, leading to estimates that MYC proteins are involved in at least 70% of all human cancers. MYC is particularly important in many therapy-resistant cancers. For example, in castration-resistant prostate cancer (CRPC), c-MYC is amplified in 45% of cases, while in late-stage neuroendocrine prostate cancer (NEPC), N-MYC is overexpressed in 40% of cases. In non-small cell lung cancer, MYC promotes resistance to targeted therapies against EGFR and ALK. In hepatocellular carcinoma (HCC), c-MYC protein is overexpressed in up to 70% of cases across all stages and grades, implicating MYC as a prevalent, early and critical event in tumorigenesis. Thus, inhibiting MYC systemically with a small molecule MYC inhibitor may lead to profound antitumor activity across many cancer types. However, to- date, no small molecule MYC inhibitor has been developed into the clinic. To address this drug target, we set out to develop small molecule MYC inhibitors suitable for development as a new therapeutic for cancer. After implementing a novel in silico screen, novel in vivo testing, extensive medicinal chemistry optimization, and thorough in vitro and in vivo characterization, we developed a novel series of direct small molecule MYC inhibitors that demonstrate profound antitumor efficacy in MYC-driven models of prostate, liver, and other tumor types. Our lead compound, MYCi606, completely eliminates HCC tumors in multiple syngeneic models and induces immunological memory in HCC, suggesting this compound has the potential to be an extremely impactful new therapeutic for liver cancer. Vortex Therapeutics has initiated IND-enabling studies on MYCi606. In this proposal, we seek funding to continue this the development of MYCi606 by completing process development and GLP toxicology studies. These efforts will position us for filing an IND in the near-term to study the first direct MYC inhibitor in human clinical trials.

NINDS - National Institute of Neurological Disorders and Stroke

PROJECT SUMMARY Epilepsy is one of the most common brain conditions affecting 70 million people globally, contributing to 5% of total disability-adjusted life-years (DALY) due to neurological disorders, and to 1.3% of all deaths, reflecting the heavy burden of the disease. Additionally, the US annual epilepsy-specific cost estimate ranges from $1,022 to $19,749 per patient, and costs are considerably higher ($138,600 per patient) for uncontrolled/treatment - resistant epilepsy. Although some approved anticonvulsant drugs are able to decrease the frequency of seizures, they do not become fully controlled and patients are generally maintained on daily multiple antiepileptic drugs to increase the efficacy of seizure control. Despite this polypharmacy approach, as many as 60% to 70% of patients continue to have seizures. Repeated uncontrolled seizures and the side effects arising from seizure medications have a negative impact on the developing and adult brain and can lead to severe impairment of neurocognitive function. KRM-II-81 is a novel γ-aminobutyric acid receptor potentiator developed by RespireRx that will improve on the anti-seizure control of standard of care (SOC) drugs by reducing side-effect burden (sedation, somnolence, cognition), enabling greater neural inhibition for seizure dampening, and reduce tolerance development and abuse liability. The efficacy and primary safety profile of KRM-II-81 has been demonstrated using pre-clinical models. In this SBIR Direct-to-Phase II project, RespireRx will develop the chemistry, manufacture and control (CMC) product synthesis method for KRM-II-81 in lab scale and scale up manufacture (Aim 1) that can be used for pre-clinical GMP toxicity evaluation. Further, the in vitro absorption, distribution, metabolism, and excretion (ADME) properties and in vivo pharmacokinetics properties of KRM-II-81 will be evaluated using cell cultures, and rat and dog models (Aim 2). In addition, pivotal IND-enabling toxicology studies will be performed in rats and dogs (Aim 3) to produce translatable data for conducting human clinical trials. These pre-clinical results along with clinical development plans will further be packaged into an IND-dossier for pre-IND meeting with the FDA (Aim 4). The successful completion of this SBIR Direct-to-Phase II project will uniquely position RespireRx to initiate Phase I clinical trials after obtaining regulatory clearance. If the broad anti-epileptic properties is confirmed in patients, without developing tolerance, KRM-II-81 would be a true breakthrough drug for the treatment of epilepsy and seizures that have become pharmacoresistant to treatment. With the unique ability to increase the quality of life by effectively treating pharmaco-resistance epilepsy, and with minimal side effects unlike other drugs currently on the market, KRM-II-81 is in line to become the gold standard in epilepsy treatment.

NCI - National Cancer Institute

PROJECT SUMMARY Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, the leading cause of cancer related deaths worldwide. FDA approval of mutant selective KRAS inhibitors (KRASi) has transformed the management of KRAS-G12C mutant NSCLC, which accounts for over 12% of cases. Yet, KRASi have major limitations. Only 30-40% of patients respond to KRASi, and many of those who do respond eventually acquire resistance. New research has identified persistent nucleoside salvage pathway activity in cancer cells as a KRASi escape route that enables their survival by providing the metabolites necessary for DNA replication and repair. Trethera is a clinical stage biopharmaceutical company that has developed a first-in-class small molecule drug, TRE-515, to block the rate limiting nucleoside salvage pathway enzyme deoxycytidine kinase (dCK). Notably, TRE-515 is the only dCK inhibitor to enter clinical development. Trethera is currently evaluating TRE-515 in a Phase 1a open- label, dose escalation study in patients with solid tumors (NCT05055609), which has enrolled 27 patients across 7 separate cohorts (40 to 720 mg). TRE-515 is well tolerated (no dose limiting clinical or laboratory toxicities have been observed in any treated patient) with mild side effects, compelling target engagement, and early signs of clinical benefit. These encouraging results highlight the immediate potential for TRE-515 to improve outcomes when administered as a component of a rationally designed combination therapy. Phase I of the proposed research (Aims 1 and 2) will establish dCK as a targetable KRASi resistance mechanism in NSCLC models. In Aim 1, the improved efficacy of KRASi with TRE-515 will be defined in two human KRAS-G12C mutant NSCLC xenograft models. In Aim 2, xenograft tumors derived from engineered dCK knockout NSCLC cells will be evaluated to confirm that blockade of cancer cell dCK activity enhances KRASi efficacy. Phase II research (Aims 3, 4, and 5) will generate the necessary data to support a clinical trial testing KRASi with TRE-515 for NSCLC. Aim 3 will apply a clinically relevant positron emission tomography (PET) imaging assay to track tumor dCK activity as a noninvasive treatment response biomarker. In NSCLC xenograft models, baseline and treatment induced alterations in tumor dCK specific PET probe uptake will be associated with combination therapy efficacy. Studies in Aim 4 will test the antitumor effects of KRASi with TRE-515 across KRAS-G12C mutant NSCLC models in vitro to determine the mechanism of anticancer activity and if specific co-occurring mutations dictate combination treatment efficacy. Therapy response will be evaluated using mass spectrometry metabolite quantification alongside flow cytometry analysis of cell cycle alterations or cell death. In Aim 5, TRE-515 will be tested with KRASi across an expanded panel of NSCLC xenograft tumor models including a KRAS-G12C mutant patient derived xenograft model and a lung orthotopic tumor model. The data generated through the successful completion of proposed Aims will (i) establish the enhanced efficacy of KRASi with TRE-515, (ii) enable a Phase 1b trial for NSCLC evaluating KRASi with TRE-515, and (iii) attract outside funding for the proposed trial.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY Alpha-ketoglutarate (aKG, also known as 2-oxoglutarate) is a key metabolic regulator of chromatin structure and, by extension, cell fate and function. aKG is imbued with this regulatory capacity based on its role as a requisite substrate for a broad class of epigenetic ‘eraser’ enzymes that catalyze the removal of methyl marks from DNA and histones. The importance of aKG in establishing and maintaining cellular identity is underscored by a series of recent discoveries that cut across normal and diseased biological states. For example, elevated aKG levels in naïve embryonic stem cells are critical for maintaining a genomic architecture that facilitates pluripotency. Conversely, many cells display aKG antagonism phenotypes that promote malignancy or alter immune cell function, which is achieved through elevated aKG catabolism or by production of metabolites that competitively inhibit aKG binding to proteins. Despite the fundamental role for aKG in establishing cellular identity, we have limited understanding of the molecular mechanisms that control the pool of aKG that is available in the nucleus to support DNA and histone demethylation. This limitation is tied to one central roadblock: a paucity of biomedical research tools that can be used to quantify changes in nuclear aKG pool size with high sensitivity and specificity. To address this issue, we have designed and collected feasibility data supporting a new biomedical research tool to measure nuclear aKG. This tool, which we term the aKG-ON biosensor system, is similar to the classical TET-ON system. Rather than responding to exogenous tetracyclines, however, the aKG-ON biosensor system responds to changes in nuclear aKG. The aKG-ON biosensor system has two components: 1) a chimeric transcription factor based on a cyanobacterial protein, NtcA, whose DNA binding activity is allosterically regulated by aKG, and 2) a synthetic reporter gene comprising a promoter with NtcA DNA binding sites and a GFP cDNA. The output of this system, GFP expression, can be used to assess levels of aKG in nuclei of live mammalian cells. Our central hypothesis is that the transcription factor NtcA presents an attractive starting point for biosensor engineering and synthetic biology given its evolutionarily conserved role in aKG sensing in cyanobacteria. We seek to develop and validate an optimized prototype of the aKG-ON biosensor system via three Aims. In Specific Aim #1, we seek to improve the dynamic range of this system through signal amplification and directed evolution approaches to produce a prototype of the aKG-ON biosensor system. In Specific Aim #2, we seek to validate utility of the aKG-ON biosensor system prototype in unbiased forward genetic studies. In Specific Aim #3, we seek to engineer the aKG-ON biosensor system into mice and validate its utility for measuring nuclear aKG levels in vivo. If successful, our work will create a new biomedical research tool that permits direct assessment of the aKG pool that is available to chromatin-modifying enzymes, thereby enabling studies of mechanisms of metabolic-epigenetic crosstalk that drive cell state transitions in development and disease.

Environmental Conservation

Development of a PER for the WWTF Upgrade Project.

NIH

Project Summary/Abstract Vein graft failure remains a major clinical challenge, with 20–30% of grafts failing within the first year due to neointimal hyperplasia and adverse tissue remodeling. Veterans, in particular, are disproportionately affected by peripheral artery disease (PAD) and coronary artery disease (CAD). Existing interventions such as drug-coated balloons (DCB) or stents and antithrombotic therapies are limited by complications related to permanent metal implants, imprecise drug dosing, and a lack of consideration for patient-specific characteristics. Moreover, no platform currently exists to screen personalized therapies prior to clinical application. To address these limitations, we propose a novel vessel-on-a-chip ex vivo perfusion model using saphenous veins harvested from patients undergoing coronary artery bypass grafting, peripheral bypass, or amputations. This model creates a physiologically relevant microenvironment for studying neointimal hyperplasia progression, predicting vein graft performance, and screening targeted therapies on a patient-specific basis. Our approach utilizes 3D-printed chips that are customized to accommodate variations in human vein size. Each vein segment is sutured into the chip, embedded in a collagen-based extracellular matrix (ECM) for mechanical support, and connected to a peristaltic pump for perfusion. The platform further incorporates pressure sensors and imageable chambers for real-time monitoring of hemodynamic parameters. Notably, each pump can operate up to eight chips simultaneously, offering a higher-throughput alternative to conventional animal models. This high-fidelity ex vivo system will enable real-time tracking of neointimal hyperplasia, correlation of pressure dynamics with disease progression, and efficient drug screening to optimize individualized treatment strategies. By bridging the gap between preclinical testing and clinical application, our platform has the potential to transform personalized vascular medicine and improve long-term graft outcomes.

NIA - National Institute on Aging

Summary Alzheimer’s disease (AD) and frontotemporal lobar degeneration (FTLD) are neurodegenerative disorders that cause dementia and represent a major public health concern. Both diseases primarily affect the elderly and are becoming increasingly prevalent as the aging population grows. These disorders are associated with pathological changes in the brain, including the deposition of abnormal proteins. TDP-43 (TAR DNA-binding protein of 43 kDa) was initially linked to FTLD and has since been found in up to 60% of brains with AD. One of the most significant challenges in understanding the role of TDP-43 in neurodegeneration and developing effective treatments for these disorders is the lack of reliable biomarkers that can predict TDP-43 pathology in living FTLD and AD patients. TDP-43 is clinicopathologically heterogeneous, and the relationships among clinical phenotypes, local neurodegeneration, and TDP-43 pathology remain unclear. This heterogeneity arises from the complex molecular composition of TDP-43 inclusions. A biomarker or antibody capable of revealing the relationship between TDP-43 pathology and neurodegeneration would significantly advance the field. Through the screening of over 5,000 monoclonal antibodies (MAbs), we identified an anti-TDP-43 antibody, MAb No. 9, which recognizes novel TDP-43 pathology in FTLD and AD. Furthermore, Meso Scale Discovery (MSD) immunoassays using this antibody detected elevated TDP-43 levels in the plasma extracellular vesicles (EVs) of FTLD and AD patients, suggesting its potential as a plasma biomarker for TDP-43 proteinopathies across different dementias. The main objectives of this project are to investigate the MAb No. 9-positive TDP-43 pathology and its association with neurodegeneration in FTLD and AD and to develop plasma biomarkers for TDP-43 proteinopathies. To achieve these objectives, we will examine how MAb No. 9-positive TDP-43 pathology contributes to neurodegeneration in FTLD and AD by analyzing the relationships between MAb No. 9 positivity, inclusion types, pathology subtypes, and local neurodegeneration in brain specimens from autopsy- confirmed FTLD and AD patients. We will also establish plasma biomarkers of TDP-43 proteinopathies by measuring plasma EV TDP-43 levels using MSD immunoassays in cases of neuropathologically confirmed FTLD-TDP, FTLD-tau (a subtype of FTLD), and AD, as well as in healthy aging controls. This study has significant

NCI - National Cancer Institute

Project Abstract: Brain metastasis is one of the most common sites where breast cancer metastasizes. Despite the recent significant advances in the treatment of primary tumors, the incidence of breast cancer brain metastases (BCBMs) has increased. This increase is most likely due to improved patient survival and thus time for cancer cell dissemination, the limited efficacy of primary tumor therapies in treating BCBMs, the selection of chemoresistant metastatic clones upon treatments, and the lack of drugs that penetrate the blood-brain barrier (BBB). Compared with other major metastasis sites (bones, liver, and lungs) where the quality of life (QofL) is still manageable to some extent, patients with BCBMs have much more severe impairments in QofL and significantly lower survival. Therefore, there is a significant unmet medical need to develop an effective treatment for patients with BCBMs. Our NCI-supported research has led to the discovery and patenting of SEAK193 that: (1) has a clear mechanism of action confirmed by X-ray co-crystal structure and other experimental data; (2) has excellent metabolic stability and PK profiles; (3) is potent against drug-resistant breast tumor models in vivo; (4) has high brain penetration and shows good efficacy in an aggressive, clinically relevant taxane-refractory (TxR) BCBM TNBC PDX; and (5) shows a promising ability to potentiate anti-PD-L1 immunotherapy, which could enhance inhibition of extracranial metastases. SEAK proposes in this STTR Phase 1 to further de-risk SEAK193 as a single agent to assess its viability as a clinical candidate not only for the effective treatment of BCBMs, but also for brain metastases from other cancer types (lung, melanoma) or primary brain tumors (glioma). Aim 1. Determine the Kp,uu,brain value of SEAK193 in rats to confirm and quantify its brain penetration. SEAK193 has high brain penetration in mice. The BBB in rats more closely approximates the humans. Thus, we will confirm and quantify brain penetration of SEAK193. Milestone (1): SEAK193 has a Kp,uu,brain ≥ 0.5 in rats. Aim 2. Evaluate the efficacy of SEAK193 in two representative models of TNBC and HER2+ BCBM and assess the potential of SEAK193 benchmarked to a standard-of-care (SOC) drug, capecitabine. We will determine the efficacy of SEAK193 in the previously validated HCI-10 TNBC PDX and in the JIMT HER2+ BCBM models, which have been shown to reproducibly produce BCBMs in NSG mice. We will use capecitabine (CAPE) as the SOC reference control. Milestones (2): SEAK193 shows comparable or better efficacy vs. CAPE in suppressing BCBM growth; (3): SEAK193 therapy results in comparable or better survival ratios vs. CAPE. Aim 3. Determine the safety and comprehensive ADME profiles of SEAK193 to further de-risk SEAK193 to support future IND-enabling studies. We will first determine the safety of SEAK193 using SafetyScan47. We will also comprehensively assess its ADME properties and other safety profiles. Milestones (4): SEAK193 has negligible off-target effects; (5): SEAK193 has acceptable ADME and safety properties. Phase II plans. We will perform IND-enabling studies for SEAK193 in Phase II.

NIH

Significance to VA: Aging in older adults can result in declines in physical function, mobility, and quality of life, leading to adverse health events including increased risk of falls, institutionalization, and mortality. Older Veterans are at an increased risk of age-related mobility declines due to greater medical complexity and high rates of mobility limitations compared to non-Veterans. Gerofit is a national clinical exercise program that aims to improve Veteran mobility and health through a tailored exercise program which emphasizes traditional strength training as part of the program. While the current program improves outcomes, the changes in mobility outcomes are clinically small and may not translate to changes in health outcomes like mortality or fall risk. Power training is an alternate type of resistance training that involves performing concentric muscle actions as fast and safely as possible, resulting in greater improvements in mobility outcomes in older adults compared to strength training. The current Gerofit program does not use power training, and introducing this type of training into the program may propel changes in mobility outcomes beyond the level necessary to improve key health outcomes like fall risk. Perspectives of Gerofit stakeholders are needed to develop a power training intervention to meet Veteran and Gerofit program needs to optimize implementation. Innovation and Impact: If successful, this study will develop the first co-designed power training intervention in older adults and establish its feasibility and acceptability for Gerofit Veterans. Introducing power training into Gerofit may propel mobility outcomes to the level necessary to produce meaningful health outcomes for older Veterans who are at risk of age-related mobility declines. Specific Aims: The overall purpose of this CDA-2 proposal is to co-design a power training intervention for Veterans engaged in Gerofit (Aim 1) and to evaluate the feasibility, acceptability, and candidate efficacy outcomes of a Gerofit power training program compared to standard of care Gerofit exercise prescription using a mixed methods approach (Aim 2) in preparation for a future efficacy trial. Methodology: Aim 1 focuses on co-designing the power training intervention by gathering feedback from Veteran and exercise providers involved in the Gerofit program through qualitative interviews. Next, local Gerofit Veteran and exercise providers will participate in the development and refinement of the power training intervention through rapid prototyping, an iterative process of testing out the program, providing feedback through focus groups, and refining the program prior to the next round of testing. The power training intervention developed in Aim 1 will be tested in Aim 2 by conducting a randomized feasibility pilot study. In Aim 2, the feasibility, acceptability, and preliminary outcomes of a power training program in older Veterans participating in Gerofit will be evaluated using an explanatory sequential mixed methods approach. Veterans will be randomized into power training and standard of care groups and will participate in exercise sessions within the Gerofit program. Clinical outcomes will be collected post training and will be compared between the two groups (power training and Gerofit standard of care). Quantitative feasibility and acceptability outcomes will be collected post training in the power training group which will guide the refinement of qualitative interview guides to gain a deeper understanding of the feasibility and acceptability of this intervention within Gerofit. Path to Translation/Implementation: Our study results, along with the comprehensive training and mentorship that I receive, will support my transition to becoming an independent VA clinician-scientist with the goal of preparing a Merit submission to establish safety and efficacy of power training to enhance older Veteran mobility and mitigate the age-related mobility decline. In addition, this power training program will serve as a framework that will be adapted to fit the context of additional Gerofit programs as well as other VA programs aimed at improving mobility in older Veterans.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary/Abstract: Adolescent girls and young women (AGYW) in sub-Saharan Africa account for one in four new HIV infections globally. Antiretroviral (ARV) drugs can prevent and treat HIV, yet non-adherence among AGYW persists as a public health challenge due to biological, economic, and social barriers. High adherence measured via drug levels in biological specimens correlates with HIV treatment and prevention efficacy, and studies demonstrate that providing drug level feedback (DLF) paired with supportive counseling improves outcomes. Furthermore, AGYWs receiving oral or extended release ARVs (like monthly vaginal rings) desire regular real-time DLF, while their healthcare providers assert that DLF facilitates candid conversations about non-adherence. Additionally, DLF can assess the effectiveness of adherence-promoting interventions by providing an objective marker of drug exposure. Yet current DLF tests are slow, expensive, and centrally analyzed. To address the critical need for accessible and inexpensive HIV DLF for AGYW, we aim to develop rapid enzymatic assays to measure HIV drug levels in routine care settings like pharmacies or clinics. Our approach represents a paradigm shift in DLF because it measures ARVs based on their enzymatic activity, enabling rapid (<30 min) and inexpensive measurement. We will expand on prior work led by Dr. Olanrewaju (PI) which developed and validated the REverse transcriptase ACTivity (REACT) assay to measure tenofovir diphosphate (TFV-DP), a nucleotide reverse transcriptase inhibitor (NRTI) used in all approved oral PrEP regimens, and expanded REACT to detect the non-nucleoside reverse transcriptase inhibitors (NNRTIs). In this proposal, we will: (1) Calibrate REACT to operate at recently established TFV-DP thresholds that indicate oral PrEP efficacy among cisgender women. We will also optimize REACT in plasma to measure the NNRTIs doravirine (DOR), rilpivirine (RPV), and dapivirine (DPV) used in oral and long-acting HIV treatment and prevention regimens that are scaling up in high- burden settings. (2) Integrate the assay into the Harmony portable device (developed by Co-I Lutz) that can be deployed in routine care settings. We will freeze-dry reagents to eliminate the need for cold reagent storage, simplify assay setup, and enhance the platform’s usability. (3) Evaluate the sensitivity and specificity of REACT for DLF among AGYW using banked clinical samples. We will measure TFV-DP and FTC-TP in banked clinical samples collected from AGYW in a pharmacologic study led by Co-I Mugwanya (R01AI155086), DPV from banked samples collected in a PrEP delivery study among AGYW (R01HD108041) led by Co-I Pintye, and DOR and RPV from people living with HIV enrolled in the University of Washington Center for AIDS Research Enhanced Specimen Collection Service. Our multidisciplinary team at the University of Washington is uniquely qualified to complete this work given our expertise in enzymatic assays, point-of-care diagnostics, clinical validation, and implementation science.

NIAID - National Institute of Allergy and Infectious Diseases

Rapid Diagnostic Assays for Self-Monitoring of Acute or Rebound HIV-Infection: This program aims to support early-stage diagnostic technologies as platforms for developing simple, low-cost, rapid diagnostic assays to enable individuals to directly detect HIV-1 during the earliest stages of initial infection or to monitor viral suppression in chronic treated infection times when antibody responses are not an accurate surrogate for viral load. This contract aims to develop RT-PCR-based technology to measure HIV-1 viral load from patient-collected whole blood using a portable analyzer, single-use cartridges, and a simple user workflow designed for patients at home and non-technical staff in clinics and other decentralized settings.

NSF

The FAA reports that as of 2020, there are over 1.7 million registered drones in the U.S., and the demand for professional drone pilots is expected to grow as industries such as agriculture, construction, film, and real estate increasingly adopt drone technology for tasks like aerial photography, surveying, mapping, and inspections. The goal of this project, at Northwest College (NWC) in Northwest Wyoming, is the development of a Remotely Piloted Aircraft Systems (RPAS) certification program that will prepare students to meet workforce needs in the region. The project will infuse best practices in curriculum design, workforce development, and work-based learning, resulting in a short-term, stackable certificate. The critical thinking, communication, creativity, problem-solving, and analytical reasoning skills involved in attaining this certificate are transferable and are valuable assets in many employment settings. The proposed RPAS technician education certificate will feature a beginning, intermediate, and advanced course with a capstone project providing a paid internship/work-based learning experience. The objectives of this project include: (1) Increasing knowledge and awareness of RPAS for the internal and external NWC community; (2) Cultivating new and nurturing existing industry partnerships; (3) Delivering a nine-credit RPAS drone education technician certificate; and (4) Offering professional development to prepare educators to infuse RPAS concepts across various STEM disciplines and related programs at NWC. This project is funded by the Advanced Technological Education program that focuses on the education of technicians for the advanced-technology fields that drive the Nation's economy. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIDCR - National Institute of Dental and Craniofacial Research

PROJECT SUMMARY (FROM ORIGINAL APPLICATION) Human papillomavirus (HPV)-driven oropharyngeal squamous cell cancer (OPSCC) has been rising in incidence in the US and worldwide over the last four decades, particularly among young men. People with human immunodeficiency virus (HIV) have an increased HPV-OPSCC risk as compared to HIV-negative individuals. However, we have a very poor understanding of the mechanisms of persistent oral HPV infections and their synergy with HIV co-infection, as well as the role of HIV in HPV-OPSCC induction and pathogenesis. The inability to histologically characterize the early stages of HPV-induced OP tumorigenesis continues to hinder efforts to diagnose OP precancerous lesions and assess cancer risk. Currently, there is no animal model for HPV-induced oral infections and malignancies in the context of HIV-induced immunosuppression. In previous collaborative efforts, the Ozbun and Traina-Dorge labs have successfully modeled cervical HPV infections and neoplasia in female rhesus macaques (RMs) using Macca mulatta papillomavirus type 1 (MmPV1), a close relative of HPV16, which causes 90-97% of HPV-OPSCC. In this R21 proposal, we will build on our prior work and collective expertise with MmPV1, simian immunodeficiency virus (SIV), and non-human primate immunology to establish a Rhesus macaque model of persistent oral HPV and HIV co-infection with which to study oral MmPV1 infections, persistence, precancer, and OPSCC induction. This model is innovative and advantageous because both MmPV1 and SIV recapitulate their respective infections and pathogenesis in RMs like their human counterparts: MmPV1 is carcinogenic in its natural RM host, and these animals develop immunosuppression following SIV infection. As HPV-OPSCCs are more common in males, we will expose immunosuppressed SIV-infected male RMs to high-titer infectious MmPV1 virions in the tongue base, the palatine tonsil, and the soft palate. In AIM 1, we will determine the oral infectivity and persistence of MmPV1 virions in SIV-infected and immunosuppressed RMs. At timed intervals and in post-mortem tissues, we will assess MmPV1 viral loads and DNA persistence at the sites of infection, and we will screen for localized viral shedding, preneoplastic oropharyngeal MmPV1 lesions, and disease progression. We will also measure circulating tumor MmPV1 DNA over time as a potential biomarker of the onset of preneoplasia and/or OPSCC. In AIM 2, we will evaluate SIV- and MmPV1-induced changes in RM biospecimens. At timed intervals, we will quantify plasma and saliva cytokine/chemokine levels by Mesoscale immunoassays and assess HPV-disease-associated markers in cytobrush exfoliated OP cells and endpoint post-mortem tissues by immunohistochemistry. These assays will identify biological pathways altered by both viruses and determine potential disease correlates. Establishing a non-human primate model for OP PV infections and precancers will facilitate expanded follow-up studies aimed at revealing mechanisms of MmPV1 infection, persistence, and progression from precancer to malignancy in the context of SIV-induced immunosuppression; it will also promote efforts to diagnose and treat OP precancers.

NIAID - National Institute of Allergy and Infectious Diseases

To develop the HIV-eCaDI device we are developing for our NIAID project R61 AI181052 “Development of a robust HIV-1 diagnostic system (HIV-eCaDI) for at-home testing” we prepare, quantify and validate HIV virions for testing as well as test that our HIV sample inactivation is effective to maintain biosafety during testing. We utilize a combination of chemiluminescent cellular cytopathology assessment for HIV-induced cell killing and absorbance-based ELISAs for p24 antigen quantification and are developing a fluorescence polarization assay for intact virion quantification. The 16-year-old Perkin-Elmer Victor X5 multimode platereader that we use heavily in our BSL-2 laboratory to perform each of these assays recently had a critical control board failure and replacement parts are no longer commercially available, which makes performance of these necessary HIV assessment assays impossible. The objective of this supplement is to correct this limitation by replacing our Victor X5 platereader with an equivalent modern Victor Nivo platereader so we can continue to quantify and validate high-quality HIV samples necessary for quantitative evaluation of the HIV-eCaDI devices.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY Pediatric ventricular assist devices (VADs) are urgently needed to support children with heart failure awaiting transplant, especially given the scarcity of donor hearts and the complications associated with congenital heart disease. Currently, no VADs are approved for long-term use in the U.S. for children weighing approximately 10–35 kg. The immediate goal of this research is to test and refine a small and durable centrifugal blood pump (named PSU Child VAD) that we recently developed for use in children approximately 10-35 kg (BSA 0.5-1.2 m2, 1-11 years of age, and mean cardiac output of 1.5-3.5 LPM) with the weight variation of ~±1kg for 1 year old and ~±10kg for 11 year old based on growth charts developed by the Centers for Disease Control and Prevention (CDC). The device will be equipped with the optional near-physiologic pulsatility. A chronic in-vivo study was conducted on a lamb for 26 days elective, demonstrating promising biocompatibility with no evidence of end-organ failure. The goal will be achieved by three complementary, but independent aims; Aim 1) to test the prototype (Rev 0.0) PSU Child VAD through a series of 30-day in-vivo studies to validate the design and determine if the system meets the design requirements. We hypothesize that the in-vivo studies with PSU Child VAD will result in hemo-/biocompatible outcomes in terms of minimal blood trauma and no end- organ failure; Aim 2) to refine the design of PSU Child VAD (Rev 1.0, 2.0) based on the outcomes from in- silico, in-vitro, and in-vivo studies by following the FDA Design Controls approach. We hypothesize that the FDA Design Control Approach-based design iteration will result in superior hydraulic efficiency and hemo- /biocompatibility. This design is expected to minimize the occurrence of thrombosis, von Willebrand factor degradation, and hemolysis while maximizing hydraulic efficiency.; Aim 3) to develop a controller capable of providing synchronous co-pulsation to provide physiologic pulse pressures. The rationale in Aim 3 is that continuous flow blood pumps operating at a fixed speed cause static, non-physiologic pressure waveforms, which is known to cause physiologic dysfunction, such as gastrointestinal bleeding, reduced baroreflex response, and destruction of von Willebrand factor (vWF). We hypothesize that the controller will be capable of modulating the pump speed to provide physiologic arterial pulse pressure, thus balancing the physiologic levels of vWF. The proposed work is innovative because it is specifically designed to provide long-term (bridge-to-transplantation, BTT) cardiac support that fits the complex anatomy of children while also accommodating their somatic growth. This is significant because it can lead to hospital discharge and avoid the need for multiple surgeries. The long-term goal of this research is to develop a fully implantable system with the ultimate aim of enhancing the survival and quality of life for children in BTT. If successful, the device will directly benefit children awaiting donor hearts by significantly enhancing both their chances of survival and overall quality of life.

DECD

Development of a strategic plan for the Town to include recommendations from the New Milford Plan of Conservation and Development

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY Current genome editing technologies, such as CRISPR nucleases, are effective for gene knock-outs but face limitations in the precise and efficient insertion of large DNA fragments into specific genomic sites, particularly in non-dividing cells. These limitations arise from the reliance on double-strand breaks and error-prone host repair mechanisms, which often lead to unintended mutations and off-target effects. Additionally, the inefficiency of homology-directed repair further constrains the potential of these methods for gene therapies requiring stable and accurate gene insertion. Consequently, a more efficient, safe, and precise system is needed. This project seeks to address these challenges by developing a platform technology based on the piggyBac (pB) transposase, which facilitates precise DNA insertion with minimal off-target effects. The approach incorporates rational design and directed evolution to optimize the transposase, enabling the delivery of large transgenes to specific genomic sequences. Unlike traditional gene editing methods, this technology actively integrates large DNA fragments without relying on double-strand breaks or homology-directed repair, making it a versatile tool for both therapeutic and research applications. Preliminary data indicate that the pB transposase, when modified with custom DNA-binding domains, achieves targeted DNA integration. Key mutations in the transposase’s native DNA-binding domain have significantly reduced off-target insertions while preserving its ability to efficiently deliver transgenes to specific genomic sites. This proposal aims to further refine the system through phage- assisted continuous evolution (PACE) to enhance both the efficiency and specificity of the transposase, establishing a highly effective gene therapy platform. The project will optimize the pB transposase for site-specific integration by tethering it to custom DNA-binding proteins. Structural modeling will guide these modifications, reducing off-target effects and enhancing targeted insertion into the human genome. PACE will be applied to evolve pB transposase variants with improved efficiency and specificity, rapidly scanning large sequence spaces to identify variants that are suitable for therapeutic applications. The clinical potential of this technology will be demonstrated by using the evolved pB transposase to deliver a chimeric antigen receptor (CAR) gene to human T-cells, highlighting its ability to efficiently and precisely deliver therapeutic genes. This platform technology represents a significant advancement over existing genome editing methods by enabling the precise, efficient, and safe integration of large DNA fragments into non-dividing cells. It offers broad applications in the treatment of genetic diseases and biomedical research, where the ability to integrate large or multiple genes at specific genomic locations is essential. By addressing the limitations of current methods, this approach has the potential to revolutionize gene therapy, providing a robust and versatile tool for a wide range of genetic interventions.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY/ABSTRACT The Mammarenavirus genus of the Arenaviridae family, encompasses several severe human pathogens, including Old World arenaviruses (OWAs) like Lassa virus and New World arenaviruses (NWAs) such as Machupo and Junín viruses, each of which cause hemorrhagic fever. Currently, the only available vaccine, the live-attenuated Candid #1 for JUNV, is not FDA-approved and provides limited neutralization breadth, leaving populations vulnerable to diverse arenaviruses. Monoclonal antibody (mAb) therapies represent one of the most important treatment opportunities in modern medicine. They are being used to treat and cure numerous diseases, including different types of cancer and autoimmune diseases. Since the 1980s, mAbs like Synagis against RSV, and Ebanga™ and Inmazeb™ against Ebola virus disease have been used to treat viral health threats. However, there remains no licensed therapies for treatment of any arenavirus infection. As such, development, especially of broadly applicable therapeutics, is an urgent need. Pioneering discoveries from our group illuminated the “Achilles Heels” on the Lassa virus surface glycoprotein and led to a first-in-class pre- clinical therapeutic for Lassa virus infection. Here, we extend this cutting-edge work to New World arenaviruses. Employing a novel panel of prefusion-stabilized NWA GPCs, we will elucidate the structural and functional properties of these proteins, providing essential templates to interpret antibody responses. Our approach includes advanced methods such as high-resolution cryo-electron microscopy to visualize nAb interactions at a molecular level. We will isolate a diverse array of broadly protective nAbs from immunized animal models and humans vaccinated with Candid #1, assessing their breadth and potency to identify candidates for therapeutic development. Furthermore, we will conduct comprehensive structural and biochemical analyses to delineate the antigenic landscape of NWA GPCs and establish mechanistic rules underlying their neutralization. This multifaceted investigation promises to yield groundbreaking insights into the immune response to NWAs, paving the way for the development of effective vaccines and immunotherapeutics to combat these unpredictable and often fatal infections.

NHLBI - National Heart Lung and Blood Institute

“Development of a tissue-targeted non-thrombotic EPO derivative.” Chronic obstructive pulmonary disease (COPD) affects 16 million people in the US and is the third leading cause of death in the world, with an estimated 500 million cases worldwide. In the US the primary cause is cigarette smoking, but in most of the world the cause is air pollution. In this disease, the airways (bronchioles) are constricted and the alveoli are distended and never fully contract. Current treatments use drug inhalers that open up the airways, but these treatments are only partially effective. In addition, about 2 million of the US COPD patients are also anemic, so that oxygen delivery is particularly poor. Erythropoietin (EPO) is a hormone that mediates the body’s response to massive blood loss (hemorrhage). EPO is used to treat anemia in kidney failure, cancer, etc., based on its stimulation of red blood cell production. EPO also has a useful tissue-protective activity that can reduce effects of oxygen limitation that damage the heart, brain, and other tissues. However, EPO also enhances blood clotting and increases frequency of heart attacks, strokes and deep vein thrombosis when given to anemic patients. To address COPD and other problems of oxygen delivery, General Biologics is developing an engineered protein, termed “EPO-H” (EPO for Hypoxia), that retains the red blood cell producing and tissue-protective activities of EPO, but lacks the blood clotting side effects. EPO-H will also have a long plasma half-life, to allow for infrequent dosing, patient convenience, and reduced burden on the health care system. The net effect should be that, compared to current commercial versions of EPO, our protein (“EPO-H”) will maintain production of red blood cells, show increase neuroprotection, and have significantly reduced or eliminated blood-clotting site effects. Natural EPO is not currently given to COPD patients or patients with disorders such as cystic fibrosis or extreme Covid-19 requiring hospitalization and mechanical ventilation. Our experimental aims are: (1) General Biologics will produce engineered protein for all of the experiments and will test the proteins for blood-based surrogate markers of hypoxia resistance and blood-clotting, to establish a dose at which the therapeutic effect is expected but side effects are not.; (2) University of Maryland will demonstrate that EPO-H can promote survival of mice in a low- oxygen environment; and also (3) demonstrate that EPO-H does not enhance clot size in a mouse model of deep vein thrombosis, even though natural EPO does increase the size of blood clots in mice with deep vein thrombosis.

NIAID - National Institute of Allergy and Infectious Diseases

To support the advanced development of candidate vaccines. This contract may support formulation and manufacture of the individual vaccine components, as well as stability testing, nonclinical immunogenicity and efficacy testing in animal models, IND enabling GLP toxicology, submission of an IND and clinical safety and efficacy evaluation.

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary: Charcot-Marie-Tooth (CMT) is one of the most common inherited neurological disorders affecting 1 in 2,500 people in the U.S. and nearly 3 million people worldwide. There are five main types of CMT with each type having numerous subtypes, such as CMT type 2E (CMT2E), and more than 100 different disease-causing genes have been identified. While most types of CMT are not lethal, these diseases significantly impact the quality of life of individuals with CMT and their families. To address CMT disease, several animal models associated with distinct genetic mutations have been developed that faithfully mimic CMT pathology providing tremendous value to our understanding of CMT disease initiation and progression. Despite these advances, no cures for any CMT subtype have been approved. We are proposing to develop a precision medicine-based approach to treating CMT2E, using a specific patient-derived mutation as the pre-clinical model developed in the Lorson laboratory. CMT2E is caused by mutations in the neurofilament light gene (NEFL). NEFL codes for the neurofilament light protein (NF-L), which, along with other intermediate filaments, including neurofilament middle and heavy, are involved in maintaining structural stability of neurons among other roles. Neurofilament aggregation is also a pathology of several neurodegenerative diseases. CMT2E-causing missense mutations have been identified throughout the various functional domains within the NF-L protein. To address the extent of NEFL mutations, we developed a mutation agnostic therapeutic approach; therefore, our therapeutic vector should address disease associated with most NEFL mutations. We developed a novel CMT2E animal model that represents a specific human patient mutation (NEFL- E396K) within the mouse Nefl gene (E397K). The Nefl-E397K mouse model faithfully recapitulates CMT2E disease. Using this disease context, we will leverage the AAV9 gene therapy system to deliver a dual cargo therapeutic vector developed in the Lorson laboratory. This vector delivers two complementary therapeutic “payloads” designed to reduce mutant NEFL, restore healthy/functional NF-L protein and prevent disease development. This project is a collaboration between several labs that bring together ideally suited areas of expertise: 1) the Lorson lab which has a long-standing interest in developing therapeutics and viral vectors for SMA, SMARD1 and other neurodegenerative diseases; 2) the Arnold lab which brings CMT experience in pre- clinical models as well as within the clinic; and 3) Dr. Hong An, an expert in bioinformatic analysis. This is a project focused upon furthering our understanding of CMT2E disease progression as well as further optimizing and validating a therapeutic for this important disease. This cross-disciplinary team is well positioned to successfully complete this project.

NCI - National Cancer Institute

Abstract Effective treatment of aggressive and treatment-resistant cancers, such as head and neck squamous cell carcinoma (HNSCC), demands innovative methodologies that accurately capture the complexities of the tumor microenvironment and predict therapeutic responses. However, traditional imaging techniques alone may not fully capture the complex and dynamic processes of tumor growth, angiogenesis, and therapeutic responses, limiting our ability to develop and optimize effective treatments. This project aims to develop a Virtual Preclinical CT (VPCT) platform to serve as a "digital twin" for studying preclinical cancer models of HNSCC. By integrating advanced photon-counting CT (PCCT) imaging with sophisticated computational models, the VPCT platform will simulate tumor behavior, optimize imaging protocols, and evaluate therapeutic interventions in a virtual setting. The VPCT platform will combine high-resolution tumor and vascular models with PCCT simulations to accurately replicate the anatomical and physiological complexity of mouse cancer models, thereby enabling precise simulations of therapeutic outcomes. Our approach is organized into three specific aims: Aim 1 focuses on developing the VPCT imaging platform by enhancing digital mouse phantoms with detailed vascular and tumor models derived from high-resolution PCCT data and integrating multiscale tumor models using computational platforms like CompuCell3D. Aim 2 will validate the VPCT platform by comparing simulated imaging data with empirical data from custom-designed phantoms and in vivo PCCT imaging of HNSCC tumors, ensuring the platform’s accuracy in material separation and vascular mapping. Aim 3 will leverage the validated VPCT platform to optimize and evaluate radiation therapy (RT) and combination treatments for HNSCC. Specifically, we will identify the optimal timing for injecting high atomic number (Z) barium nanoparticles (VivoVist™) in relation to RT to maximize vascular disruption and enhance the enhanced permeability and retention (EPR) effect. Additionally, we will explore the synergistic potential of combining VivoVist™ nanoparticles with RT and the chemotherapy agent Doxil (liposomal doxorubicin). By integrating advanced radiomics, machine learning, and computational pathology, we aim to understand therapeutic strategies and significantly improve the accuracy of treatment outcome predictions. The VPCT platform is anticipated to significantly improve tumor modeling and optimize PCCT protocols for assessing the effects of RT on tumor vasculature and evaluating combination therapies. By focusing on vascular changes and the synergistic effects of combined treatments, the platform will offer critical insights for refining RT and chemotherapeutic strategies in cancer treatment. Additionally, integrating advanced computational techniques will enhance predictive accuracy, enabling preclinical findings to directly inform clinical applications and advance personalized cancer therapies.

DEEP

Development of a walkway along a portion of the Steele Brook Stream corridor