NCATS - National Center for Advancing Translational Sciences Grants

NCATS - National Center for Advancing Translational Sciences

Anti-sense oligonucleotide (ASO) drugs have shown great potential in the treatment of human diseases. However, the lack of therapeutic efficacy and intolerance of adverse drug reactions (ADRs) and toxicity have led to the termination of numerous ASO candidates in the development pipelines, as well as the discontinuation from the market after approval by the FDA. Hepatotoxicity and nephrotoxicity are two major concerns for the FDA-approved ASO drugs in the market as well as development of new ASO-based therapies. Significant knowledge gaps still exist which prevent us from better understanding the lack of effectiveness and existence of ADRs and toxicity. One of these gaps lies in how an ASO drug goes through absorption, distribution, metabolism, and excretion (ADME) in its target organs and cells. Both in vitro cellular models and in vivo animal models have been used to assess ADME and toxicity of ASO drugs; however, they have numerous limitations. While cellular models lack human complexity, animal models lack human-specificity. Microphysiological systems (MPSs) may provide a solution to overcome this unmet need by providing human specificity and complexity as an in vitro platform. Here, we propose using the Javelin Liver Tissue Chip Plus (LTC+) platform, which integrates human liver and kidney MPSs, to study distribution, metabolism, excretion, and toxicity profiles (hepato- and nephrotoxicity) of four FDA-approved ASO drugs.

NCATS - National Center for Advancing Translational Sciences

Abstract Support is requested for a Keystone Symposia conference entitled “iPSCs: Progress, Opportunities, and Challenges,” organized by Drs. Shinya Yamanaka, Yanhong Shi and Yasushi Kajii, with scientific programming input from Keystone Symposia. The meeting will take place January 26–29, 2026 at the International Conference Center (ICC) Kyoto in Kyoto, Japan. This conference is being held to mark the 20th anniversary of breakthrough discoveries in induced pluripotent stem cell (iPSC) technologies, which have matured into viable platforms for embryology and disease modeling, drug discovery, and cell-based therapy development for a variety of human diseases. Moreover, the combination of iPSC technology with three-dimensional organoids, organ-on-chip and the emerging technologies of AI and machine learning ensures that iPSC-based platforms will yield new applications in biomedical and translational science. These innovative technologies and their applications are the primary focus of this meeting. Therefore, this conference has been designed to gather the leading scientists from academia and industry to push forward basic knowledge and medical application of iPSCs, especially those being tested in clinical trials worldwide, toward new insights and potential drugs. Additionally, this Keystone Symposia conference will provide a unique opportunity for researchers, clinicians, industrial experts and investors to interact, creating unusual collaborative prospects. Moreover, this conference will provide a rare opportunity for attendees to hear from renowned stem cell researcher and Nobel Laureate, Dr. Shinya Yamanaka, who will be giving the Keynote Address and is one of the meeting organizers. The sharing of knowledge at this meeting is expected to be transformative for the field and lead to the development of new cellular platforms and therapeutic products, which will ultimately impact clinical practice favorably.

NCATS - National Center for Advancing Translational Sciences

PROJECT SUMMARY/ABSTRACT The objective of this proposal is to define the molecular mechanisms and identify new therapeutic strategies for of an understudied class of myopathies, specifically X-linked myopathy with excessive autophagy (XMEA). XMEA is characterized by elevated levels of autophagy due to disruptions in the autolysosome function. One MEA of interest is X-linked myopathy with excessive autophagy (XMEA), a rare autophagic vacuolar myopathy that characterized by progressive proximal muscle weakness, high levels of serum creatine kinase and accumulation of autophagic vacuoles. XMEA is caused by pathogenic mutations in the VMA21 gene in which N- terminal loss-of-function variants result in early death by 10 years and milder pathogenic VMA21 splicing variants result in a slower disease progression. Patients with VMA21 pathogenic mutations have a defective autophagy and an impaired ability to form the autophagosomes. VMA21 is a subunit of the V-ATPase protein pump and its disruption results in a failure to properly acidify the autolysosome resulting in the formation of vacuolar inclusions in XMEA. No extensive biomarker studies have been performed in the XMEA population resulting in a dearth of knowledge and the lack of suitable XMEA models is a significant barrier towards any effective treatment. We have generated a Vma21 knock-in (Vma21 KI) mouse model based on an RNA-splice mutation identified in a set of XMEA patients observed at our Children’s of Alabama muscular dystrophy clinic. Vma21 KI mice have a progressive muscle weakness, impaired muscle function, and have vacuolar inclusions that form as they age, which phenocopies the XMEA patient symptoms. In parallel, we generated vma21 mutant zebrafish that have a severe loss-of-function (LoF) pathology resulting in muscle paralysis, vacuolar inclusion bodies, and early lethality by 10 days post fertilization (dpf). An autophagy drug library screen of our vma21 mutant zebrafish identified edaravone, an FDA-approved autophagy and oxidative stress inhibitor for ALS, as the most corrective compound out of 29 leads for XMEA zebrafish pathologies. This proposal seeks to establish molecular and therapeutic biomarkers for XMEA based on our analysis of XMEA patient cells, and VMA21-defective zebrafish and mouse models, with an emphasis on the Vma21 KI mice. Proteomic evaluation of the muscles from Vma21 KI mice will allow us to identify VMA21-dependent factors that progress with XMEA disease status. We also seek to evaluate the therapeutic mechanism of action for edaravone in a 6 month treatment of our Vma21 KI mice. These studies seek to establish the XMEA/VMA21 disease processes while advancing a promising autophagy inhibitor compound to eventually treat these XMEA patients suffering from this devastating neuromuscular disorder.

NCATS - National Center for Advancing Translational Sciences

Project Summary/Abstract Community pharmacists are playing a growing role in improving public health and increasing access to preventive care via pharmacist-provided public health services beyond traditional dispensing and medication counseling.1-4 Pharmacists in various states are allowed to prescribe contraceptives, PEP/PrEP for HIV prevention, tobacco cessation products, test and treat for minor ailments, and more.1-4 By providing these services, pharmacists have demonstrated their ability to support public health, decrease chronic diseases, increase patient access to preventive care, and reduce health care costs.5-10 However, implementation of these public health services in community pharmacies remains low, despite widespread interest from pharmacists and patients.11-26 The current gap is that while a lack of innovative training strategies that reflect current pharmacy practice environments has been identified as a major barrier to implementation, no known solution exists. Our solution, the Preparing Pharmacists for Public health Services (PrePPS) software will be available to pharmacists for continuing education credits and pharmacy students through pharmacy school curricula. Our business, EmpoweRx, Inc has experience developing software for pharmacists to address barriers to implementation of pharmacist-prescribed contraceptive services as a part of an SBIR Phase II award from the Centers for Disease Control and Prevention. We will use this expertise to inform our work to support development of a training software to prepare the current and future pharmacist workforce and support implementation. In Phase I of this grant, we will gain an understanding of end-user (pharmacists and pharmacy student) needs when it comes to preparing pharmacists for providing public health services as we work through the first five iterative phases of the Design Thinking process: 1) Empathize, 2) Define, 3) Ideate, 4) Prototype, and 5) Test.27 In Phase II, we will complete the final phase of the Design Thinking process: Implement. The contribution of the proposed research is a market-ready PrePPS software that will increase pharmacists' and pharmacy students’ readiness and implementation of pharmacist-provided public health services. At the conclusion of this project, we will have a market-ready, proven, user-informed PrePPS software application that will facilitate future and current pharmacist workforce readiness to provide public health services and the implementation of pharmacist-provided public health services.

NCATS - National Center for Advancing Translational Sciences

ABSTRACT This proposal addresses the need to investigate understudied proteins associated with rare diseases, such as Brugada Syndrome (PAR-25-122). One class of proteins highlighted in this RFA—Scn2b, Scn3b, and Scn4b— belongs to a family of β subunits that associate with the large pore-forming α subunits of voltage-gated sodium channels (NaV), which regulate electrical excitability throughout the body. In total, there are four distinct β subunits that can mix and match with nine different α subunits. Beta subunits are widely recognized for their ability to regulate the gating properties, trafficking, and pharmacology of Nav channel complexes. Dysfunction of these subunits has been linked to several human diseases, including epilepsy and cardiac arrhythmias such as long QT syndrome, atrial fibrillation, and Brugada syndrome. Additionally, mutations in NaV α subunits have been implicated in rare diseases, including SCN8A encephalopathy (SCN8A/ NaV 1.6), hereditary sensory and autonomic neuropathy type 7 (SCN11A/ NaV1.9), and dilated cardiomyopathy-1E (SCN5A/ NaV1.5). A critical step in understanding how beta subunits contribute to disease is elucidating their precise modulatory effects on NaV function. Electrophysiological studies in heterologous cells have demonstrated the ability of beta subunits to influence channel gating, pharmacology, and trafficking. However, results across multiple studies have been inconsistent, often due to variability in the cell lines used. A major confounding factor is that many cell lines endogenously express beta subunits, which can interfere with exogenously introduced β subunits under investigation. To overcome this limitation, we developed a specialized cell line lacking all β subunits, including Scn2b, Scn3b, and Scn4b, as well as Scn1b, MPZ, MPZL1, MPZL2, and MPZL3. These cells, termed beHAPe cells (beta- eliminated haploid cells for expression), provide a controlled system to study NaV channel regulation. Our initial electrophysiological studies using beHAPe cells reveal novel properties of beta subunits in modulating NaV1.5, the primary α subunit in cardiac tissue. Building on these findings, we propose to produce stably-expressing human (HEK) cell lines to systematically define the roles of Scn2b, Scn3b, and Scn4b in modulating additional α subunits, including NaV1.6 (a key subunit in the central nervous system) and NaV1.7, NaV 1.8, and NaV 1.9 (which are predominant in the peripheral nervous system). This work will provide deeper insights into their function in these tissues and their associated diseases. Additionally, our new data suggest that Nav1.8 plays a previously unrecognized role in cardiac function alongside NaV1.5. Understanding how β subunits modulate pore-forming subunits could provide new insights into their involvement in cardiac arrhythmias, expanding their known roles beyond the nervous system. Taken together, in addition to providing new information on the understudied Scn2b, Scn3b, and Scn4b proteins, our newly generated stable cell lines will enable studies for the development of novel therapeutics for isoform specific modulation of specific α- and β-subunit pairs.

NCATS - National Center for Advancing Translational Sciences

The Yale Center for Clinical Investigation (YCCI) was created in 2005 to advance Yale's clinical research mission. One year later, YCCI became the home of the Yale CTSA. At YCCIs inception, Yale was a national leader in T0-T2 translational research, basic/translational science training, and it supported distinctive translational science T3-T4 fellowship programs such as the Robert Wood Johnson Clinical Scholars Program. Since then, the CTSA has had a transformative impact linking all components of the Yale community in T1-T4 research, providing the central infrastructure for the effective conduct of ethical, innovative, rigorous, and reproducible research, and in training the next generation of research leaders. By any metric of scale, breadth, quality, and impact, both the CTSA's research enterprise and its educational mission have been enormously successful for Yale. This renewal application does not simply seek to maintain excellence, but to enable YCCI to drive the continued transformation of the Yale T1-T4 translational research mission and its predoctoral and postdoctoral training mission and to promote collaboration across CTSA hubs. First, it will support informatics and computational advances that drive the emergence of a learning health system. In so doing, it will draw on the Yale New Haven Health System, a six-hospital 2,681-bed consortium that provides more than 2.4 million outpatient visits annually from patients from upper Westchester county, throughout Connecticut, and southern Rhode Island. It will also prepare young scientists to draw on this infrastructure to conduct research that influences the future of healthcare. Second, it will support technological and scientific advances in areas that will support the emergence of personalized healthcare, including multi-omics and imaging. YCCI will provide pilot grant support and training to foster the development of research careers and research teams that can deepen our insights into pathophysiology and build toward personalized treatments. Third, it will engage a broader and multidisciplinary group of faculty, trainees, and community representatives to collaborate to improve health outcomes that constitute a major burden on patients, their families, and on public health. To support this mission, YCCI will also foster the development of careers in community-based research from a multidisciplinary group of young investigators and enhance the overall clinical research workforce.

NCATS - National Center for Advancing Translational Sciences

Project Summary/Abstract Domestic pharmaceutical manufacturing is currently struggling to meet growing demands. To address this challenge, it is crucial to develop tools that can significantly enhance the productivity of existing production facilities. Pharmaceutical production today relies heavily on batch-based chemical synthesis, where chemical reactions occur in several separate steps within large reactors. Among these steps, hydrogenation stands out as a very frequent and important chemical transformation unfortunately plagued by significant technical challenges, including scalability issues and safety risks due to the explosive nature of hydrogen. Additionally, the process requires expensive precious metal catalysts, further complicating production. We propose the development of a safe, scalable, and continuous hydrogenation solution for pharmaceutical applications. This innovative approach will combine W.L. Gore’s advanced catalyst immobilization technique with a layered modular reactor design designed by Zaiput. Our goal is to overcome the current challenges associated with hydrogenation, such as safety concerns, scalability limitations, and the inefficiencies inherent in traditional batch-based chemical synthesis. In Phase I of this SBIR project, we will focus on key technological innovations necessary to demonstrate the scalability of the layered reactor concept. Specifically, we will: 1) develop an advanced gas-liquid distribution module that ensures uniform distribution across various reactor sizes, and 2) demonstrate the system’s effectiveness by achieving scalable reaction results. In Phase II, we will design and build larger-scale devices, conduct real-world testing in partnership with pharmaceutical manufacturers to validate the technology in a manufacturing setting, and initiate the commercialization process. If successful, this project will produce a tool capable of immediately addressing hydrogenation challenges and contribute to the reshoring of pharmaceutical manufacturing to the U.S. This aligns with government priorities by offering safer, more efficient, cost-effective, and environmentally friendly hydrogenation processes.

NCATS - National Center for Advancing Translational Sciences

PROJECT SUMMARY The 2026 FASEB Summer Research Conference (SRC) on The Ubiquitin System: Mechanisms, Functions, and Therapeutics (UB) will be held June 1-4, 2026, at the Galt House Hotel in Louisville, KY. The UB SRC has been held biennially for nearly four decades, since 1989, and is the premier forum for sharing and discussing the latest developments in understanding the basic biology of processes regulated by ubiquitin and ubiquitin- like (UBL) proteins, and therapeutic approaches that target or harness these systems to treat disease. This application requests support for 12 new and early career invited speakers and 2 trainee organizers of a pre- meeting career forum to attend the 2026 SRC, which will bring together approximately 150 scientists at all career stages, from graduate students and postdoctoral fellows to principal investigators, professors, and company leaders working in academia, government, and industry. Regulated protein degradation through the ubiquitin-proteasome system is a critical aspect of protein homeostasis, the cell cycle, gene expression, metabolism, and development. Ubiquitin and UBLs also regulate a myriad of other cellular functions, including vesicular trafficking, autophagy, signal transduction, and membrane protein biogenesis, via non-degradative mechanisms. Hence, ubiquitin and UBL pathways impact essentially all aspects of eukaryotic cell biology and thus human health and disease, including developmental, neurologic, and inflammatory disorders, as well as cancers. The 2026 UB SRC will feature 8 themed sessions covering a broad range of topics focused on ubiquitin and UBL systems. These sessions will include 28 invited speakers and 24 short or “lightning” poster talks that will be selected from the submitted abstracts. In addition, for the first time, the 2026 UB SRC will co- locate and run concurrently with the FASEB SRC on Protein Folding in the Cell (PF), highlighting the close connection between protein biogenesis and degradation while also maximizing cost savings. The organizers of the two SRCs have joined forces to coordinate the programs of each SRC to maximize interactions. This includes 3 joint sessions, including the opening keynote lectures, running the poster sessions concurrently, and scheduling all coffee breaks, on-site meals, and panel and roundtable discussions to be held together. In addition, there will be a joint half-day career forum before the main SRCs, aimed at introducing trainees to each other, the SRC organizers, and invited speakers. This pre-meeting forum will be organized by a peer group of 4 trainees and provides opportunities for 16 additional trainees (8 each from UB and PF) to present their work. Finally, a new outreach component will be introduced to the SRC through the invitation of local high school students to attend the poster sessions. Thus, in addition to upholding the long-standing tradition of the UB SRC in featuring the latest high-quality work from basic science to translation to the clinic, these innovations will distinctly enhance and broaden the UB community to foster a safe, welcoming, and stimulating environment for sharing nascent ideas and unpublished results and forming new collaborations.

NCATS - National Center for Advancing Translational Sciences

SUMMARY We discovered mutations in Tudor family RNA-binding protein (RBP) TDRD7 (OMIM: 611258) cause congenital birth defect cataract–loss of eye lens transparency–in humans. We linked TDRD7 to defects in sperm formation, leading to the recognition of a rare novel human syndrome that includes congenital cataract and azoopermia as symptoms. Cataract occurs in neonates as a rare condition, but causes permanent vision damage, with surgery being the only treatment. Even after surgery, patients face eye complications throughout life. Thus, new therapies are urgently needed. Yet, knowledge on TDRD7, especially on its function in the eye, is limited. Thus, we will address this critical knowledge-gap by identifying potential new druggable pathways linked to TDRD7. Lens differentiation upregulates select RNAs/proteins while they undergo dramatic cell-shape changes–involving ~1000-fold length-wise increase–and migration toward lens core. A long-standing question is, what mechanisms control these complex cellular differentiation events? Our data suggests the involvement of TDRD7. TDRD7 protein has OST-HTH/LOTUS and Tudor domains that may allow it to associate with RNA and methylated arginine/lysine, respectively. Our data shows Tdrd7 knockout mice (Tdrd7KO) exhibit cataract and reduced expression of genes linked to human/animal lens defects. Further, Tdrd7 loss causes severe cellular morphology defects in mature lens fibers. Our data shows that in addition to abundant Tdrd7 protein in the fiber cytoplasm, where it participates in protein-RNA complexes, Tdrd7 protein also enters fiber nucleus beginning at midembryonic stages. These exciting findings lead to a paradigm-shifting hypothesis: TDRD7 may participate in both (1) post-transcriptional control and (2) chromatin control, to facilitate proper gene expression regulation in the lens. This will be tested by pursuing the following goals: Characterize spatiotemporal chromatin and transcriptome changes in Tdrd7KO mouse lens at the single-nucleus level and use AI-based approaches to derive regulatory networks (Aim 1). Characterize the impact of Tdrd7-loss on lens proteome and identify its protein interactions in normal lens (Aim 2). This innovative proposal will fundamentally advance knowledge on TDRD7 by: (1) defining, on single nucleus level, spatiotemporal changes in lens transcriptome and (2) changes in lens chromatin, upon Tdrd7 loss, (3) defining proteins impacting Tdrd7 function and those altered in Tdrd7KO, and (4) making this regulatory information publicly available via a web-based, user-friendly resource iSyTE for continued eye gene discovery. We will collaborate with Dr. Shinichiro Chuma (Kyoto University, Japan) who is an expert on TDRD-proteins and has developed a Tdrd7 knockout (KO) mouse model that we will investigate. While the facilities in US and Japan are similar, the targeted Tdrd7KO mouse model is not commercially available in the US and Dr. Chuma’s 20 years expertise on TDRD-proteins (e.g. advise on Tdrd7 biochemical protocols) is necessary for success of the aims. This translational research will advance knowledge on an understudied protein, TDRD7, and identify potential new drug targets/pathways for novel therapies/treatments for cataract birth defect.

NCATS - National Center for Advancing Translational Sciences

Project Summary Chimeric antigen receptors (CARs) redirect T cells to eliminate antigen-expressing cells by coupling extracellular antigen binding domains to intracellular T cell signaling domains via hinge and transmembrane domains. Each domain of a CAR plays a crucial role in shaping CAR T cell phenotype and function. Despite the fundamental relationship between CAR design and T cell phenotypes, efforts to optimize CAR design have been limited and low-throughput. Consequently, most CARs in the clinic utilize the same basic design architectures, likely contributing to suboptimal performance. The overall goal of this Phase I STTR project is to develop a modular, high-throughput screening platform to optimize CAR designs for any antigen, disease, and modality. The central hypothesis is that CAR design optimization can address key challenges such as tonic signaling, antigen sensitivity, and antigen loss. To achieve the overall goal, the research proposal is structured into three specific aims focused on (1) screening large libraries of CD30 and (2) BCMA CAR designs to overcome diverse and fundamental challenges, and (3) building computational models to predict optimal CAR designs. The first aim is motivated by the profound signatures of tonic signaling of a clinically tested CD30 CAR design. Previous data suggest that multiple domains of a CAR can influence tonic signaling. To identify optimal CAR designs that reduce tonic signaling and retain antigen-specific potency, a large (~105 member) library of hinge, transmembrane, and CD3z variants will be screened using readouts of dysfunction and antigen potency. The second aim seeks to showcase the modularity of the platform by addressing a different challenge – increasing antigen sensitivity and reducing antigen loss – of a commercial BCMA CAR. The BCMA CAR exhibits impaired cytokine production in vitro and tumor control in vivo in response to tumor cells with low levels of BMCA. To identify optimal CAR designs that increase antigen sensitivity and reduce antigen loss, a large (~105 member) library of hinge, transmembrane, and CD3z variants will be screened using readouts of antigen sensitivity and antigen loss. The third aim will leverage these screening datasets to develop computational models that predict CAR activity, expanding the search space from empirical data of ~105 CARs to all possible ~1011 CAR designs. This research proposal is strongly aligned with the mission of NCATS to turn research observations into health solutions through translational science. Developing a screening platform will expedite the scale and speed at which research observations of CAR designs can happen and translate into life changing therapies. In the short- term, this proposal aims to discover optimal CD30 and BCMA CAR designs to improve CAR T cell therapy for Hodgkin Lymphoma and Multiple Myeloma. This proposal establishes the foundation of a long-term goal to engineer more effective CAR T cell therapies for indications with high unmet need.

NCATS - National Center for Advancing Translational Sciences

Summary Rare diseases collectively affect approximately 30 million people in the U.S., posing a significant health and economic burden. Despite their impact, the genetic and molecular bases of many rare diseases remain poorly understood, hindering the development of effective diagnostics and therapies. This proposal focuses on elucidating the functions of a subset of understudied proteins associated with rare diseases, identified in NIH PAR-25-122, that may play critical roles in left-right (LR) patterning during embryonic development. Disruptions in LR patterning underlie heterotaxy syndrome, a rare congenital disorder characterized by mispositioned internal organs and often severe cardiac malformations. Knowledge gained on how these proteins function in LR patterning can then be leveraged to understand how the protein may function to produce rare disease phenotypes in other tissues and will identify genes that should be evaluated as causative for Heterotaxy in humans. Building on extensive expertise in zebrafish models of LR development, the project aims to (1) assess the role of candidate proteins in LR patterning using antisense and CRISPR-based approaches, (2) generate targeted mutations in genes affecting LR development, and (3) develop transgenic zebrafish lines and antibodies to enable functional studies and lay the groundwork for future drug screening efforts. Zebrafish provide a powerful in vivo system to investigate gene function and conduct high-throughput drug screens. The proposed research will advance our understanding of rare disease gene function in LR patterning and establish essential tools to support therapeutic discovery in future proposals.

NCATS - National Center for Advancing Translational Sciences

Project Summary INTS12 is a chromatin interacting subunit of the mammalian Integrator complex that binds to accessible chromatin and controls various aspects of transcription such as elongation and termination. INTS12 interacts with negative elongation factors as well as RNA Polymerase II and regulates its pausing and release into productive elongation. Studies on INTS12 biology in various mammalian systems such as hematopoietic tissues is lacking despite ample evidence of its presence and likely contributions to hematopoietic development and disease. For instance, INTS12 is highly expressed in early stage erythroid progenitor cells during ex vivo erythroid differentiation of human CD34+ hematopoietic stem cells, and it is significantly reduced in the rare hemolytic anemia Congenital Dyserythropoietic Anemia Type IV (CDA IV; CDAN4), which affects young children and renders them transfusion dependent. This proposal will address our core hypothesis that INTS12 function is important for transcription regulation during normal human red cell development and reduced INTS12 levels in CDA IV red cells contributes to ineffective erythropoiesis due to disrupted RNA Polymerase II elongation control. To achieve this, a newly established erythroid progenitor cell line called BEL-A that can be expanded indefinitely, and differentiated to mature erythroid cells will be used. Aim1 will focus on INTS12 chromatin binding in normal wild type (WT) and CDA IV mutant BEL-A cells, and this will be correlated with existing unpublished chromatin accessibility and transcription factor binding data from erythroid stage-matched WT and CDA IV BEL-A cells. Further, the effect of depleting or overexpressing INTS12 on red cell development in WT and CDA IV will also be characterized. In Aim2, we will explore INTS12 functions in transcription by assessing the impact of INTS12 perturbations such as depletion and overexpression on nascent gene expression and RNA Polymerase II occupancy. These data will be correlated with INTS12 occupancy determined from Aim1, and any alterations in nascent transcription leading to changes in RNA Pol II pausing and elongation due to perturbed INTS12 levels will suggest that INTS12 contributes to altered transcription regulation in CDA IV. Transcription elongation by RNA Polymerase II is highly regulated and involves many transcription co- factors and epigenetic mechanisms, some of which are chemotherapy targets for certain hematological malignancies. Further, INTS12 interacts with chromatin using a conserved PHD domain that is being investigated as a potential chemotherapy target using derivatives of a class of compounds known as Amiodarones. The knowledge harnessed from this proposal will thus enable future investigations into INTS12 and Integrator complex biology in erythropoiesis, as well as translational studies on the potential for INTS12 as a therapeutic target in CDA IV patients.

NCATS - National Center for Advancing Translational Sciences

SUMMARY Lysosomal storage diseases (LSDs) are a group of inherited disorders that arise from mutations in genes responsible for encoding lysosomal enzymes or transporters, which are critical for breaking down and exporting complex macromolecules. When these enzymes or transporters fail to function properly, harmful substances accumulate within cells, leading to widespread cellular dysfunction and severe clinical abnormalities. In particular, the sphingolipidoses—a subset of LSDs—cause dysfunction in the breakdown of sphingolipids, essential components of cell membranes and regulators of critical signaling pathways. This disruption not only impairs cell function but also triggers a cascade of damaging effects that can significantly diminish quality of life. Gaucher disease (GD) is the most common sphingolipidosis. It occurs due to a deficiency in the enzyme β- glucocerebrosidase (GCase), leading to the buildup of substrate glucosylceramide (GCs) in macrophages, eventually resulting in various complications. The clinical phenotype is variable, but three clinical forms have been identified: type 1 is the most common and typically causes no neurological damage, whereas types 2 and 3 are characterized by neurological impairment. Once diagnosed, GD typically requires lifetime treatment. Regulatory T cells (Tregs) have a unique ability to cross the blood-brain barrier and to secrete anti-inflammatory factors, making them a promising foundation for new cell-based therapies. Our innovative approach allows Tregs to be engineered to deliver various therapeutic molecules directly to affected tissues and disease sites, offering a promising treatment platform for protein replacement therapies. In this application, we propose to engineer Tregs to secrete and replace GCase levels in the brain and peripheral tissues through the following aims: Aim 1: Generate GCase expression constructs with and without a MOG CAR for expression and lab-scale production testing. Aim 2: Evaluate expression and functional activity of engineered Tregs in vitro. Aim 3: Demonstrate in vivo expression of GCase and preferential localization of GCase -Tregs to the brain and peripheral tissues, local production of GCase, and reduction of GCs in affected tissues. Phase I Milestone: Select lead GCase-Treg product based on its ability to be detected in the brain and peripheral tissues within seven days of injection, and its ability to produce sufficient GCase >5-fold over vector control Tregs) to significantly reduce the levels of GCs in affected tissues. The selected candidate will be developed and advanced through IND-enabling studies in Phase II, including required studies for translation of this therapy into the clinic.

NCATS - National Center for Advancing Translational Sciences

Abstract/Summary Cell free DNA (cfDNA) analysis is a translational method that has revolutionized both basic science and clinical medicine. It has allowed basic understanding of gene function to impact clinical decisions and allowed clinical specimens to be used for basic science. It has become part of the standard of care for perinatal screening, monitoring transplants, and choosing therapy for cancer. Clinical trials evaluating cfDNA in virtually every disease, including diseases of heart function, infections, vascular disease, neurology, psychiatry, organ transplantation, eclampsia, fetal health, inflammatory/autoimmune diseases, and cancer are showing promising results. Even just the easily measured level of cfDNA is a useful screening tool for maternal risk, many other tests use relatively simple PCR. cfDNA is the basis of highly sophisticated multiple cancer early detection (MCED) and potentially screening for other diseases, even including Alzheimer’s. Most applications use blood cell free DNA (BcfDNA). BcfDNA turns over in under 2 hours, it is subject to variation over the course of the day, it needs to be collected in special tubes to be stable, or immediately processed with biohazard care, to obtain plasma, which needs to be stored frozen or immediately processed by “moderate complexity” lab cfDNA isolation methods, which yield variable amounts of BcfDNA. Typically, patients’ visits are weeks or months apart and only 1 -20 mls of blood is obtained yielding 10s of nanograms of cfDNA. For MCED, MRD, CNS diseases, and occult infections, this small amount limits sensitivity. Urine also contains cfDNA (ucfDNA), at comparable or a bit lower concentration. Urine can be collected multiple times a day and on as many days as desired at home without the cost, inconvenience and potential exposure to infectious diseases, of the often immunosuppressed patients, inherent in a clinic visit. However, existing methods to extract ucfDNA only allow small amounts of urine to be processed; they too are complex methods that have variable, generally low, efficiency. When scaled up they are prohibitively expensive. Even with preservatives, there is immediate degradation of a fraction of the DNA before the specimens reach the lab. Numerous studies show ucfDNA contains cfDNA from the blood and that this trans-renal DNA can be used to understand system functions. Included preliminary data document a use anywhere, all resource sites (even home use), simplified and inexpensive method collects 20x to 100x more cfDNA, immediately separates the cfDNA from enzymes that cause degradation by affinity binding, and allow transport, without biofluid and cold chain concerns. Each of the underlined items are in themselves novel. The aims are to confirm and extend this data to commercialize a system, which once received in the lab can generate cfDNA suitable for PCR and next generation sequencing in 10 minutes. ucfDNA collected over time complements the snapshot BcfDNA provides. Larger total amounts of cfDNA improve sensitivity and provide excess material so this clinical resource will be more available for basic research. While BcfDNA is a limited resource, ucfDNA is virtually unlimited. Such translational research is expected to continue to rapidly change clinical practice. Other methods are for collection or lab isolation, uProcess does both, less expensively and in 1/10 the lab time. Most importantly, the variation featured herein selects for the trans-renal DNA indicative of systemic disease; all current kits yield >80% urothelial DNA.

NCATS - National Center for Advancing Translational Sciences

Adeno-associated viruses (AAVs) are the leading vectors for gene therapy, valued for their safety, low immunogenicity, and broad tissue tropism. However, large-scale clinical translation remains hindered by challenges in manufacturing quality control. A critical barrier is the inability to rapidly and reliably measure capsid load and the ratio of full to empty capsids—key attributes that directly affect therapeutic potency and safety. Current assays such as ELISA (capsid titer), ddPCR (genome titer), and AUC (full/empty ratio) are costly, labor-intensive, and serotype-dependent, leading to inconsistent results across laboratories and production runs. To overcome these limitations, we will develop NANO-PAVE (NANOpore-based analysis of Proportion of full and empty Adeno-associated Virus capsids via nanopore Evaluation), a solid-state nanopore platform that analyzes individual AAV particles without labels, dyes, or antibodies. By leveraging voltage-driven translocation and electro-deformation, NANO-PAVE directly links mechanical signatures to genome packaging state, enabling real-time, quantitative discrimination between full and empty capsids. In addition, its single-particle resolution will allow accurate measurement of capsid load across serotypes, providing a powerful complement to existing bulk assays. This approach is cost-effective, scalable, and adaptable to all serotypes, making it ideally suited for integration into manufacturing pipelines. Phase I will establish feasibility by demonstrating robust particle discrimination, validating nanopore- derived measurements against orthogonal benchmarks, exploring genome-structure effects on capsid mechanics, and testing multiple AAV serotypes to ensure broad applicability of NANO-PAVE across vector types. Phase I deliverables will include a validated serotype-independent NANO-PAVE technology—integrating optimized nanopore design, recapture protocol, and machine-learning analysis—benchmarked against established assays to provide a rigorous, quantitative QC approach. This work will lay the foundation for Phase II development of a scalable, in-process analytical tool capable of distinguishing therapeutically relevant capsids from product-related impurities (empty, partially filled capsids, aggregates) and process-related impurities such as host cell proteins and media components, enabling comprehensive quality assessment. Ultimately, NANO-PAVE has the potential to become a transformative QC platform for academic, clinical, and industrial gene therapy programs, improving safety, efficacy, and manufacturing efficiency.

NCATS - National Center for Advancing Translational Sciences

ABSTRACT Gastrointestinal (GI) motility disorders such as irritable bowel syndrome and inflammatory bowel disease impose substantial medical and economic burdens, yet drug development remains hampered by preclinical models that fail to recapitulate human physiology and pathophysiology. Our long-term goal is to enable mechanism-informed discovery of safe and effective therapeutics by developing novel human models that accurately reflect human biology. The overall objective of this Phase I SBIR is to establish, validate, and deploy a scalable human peristaltic intestinal organoid (peristaltic-HIO) platform that couples quantitative functional readouts with deep molecular and cellular profiling. Our strong preliminary data shows self-organized peristaltic activity in pluripotent stem cell–derived HIOs containing the enteric nerons, smooth muscle, interstitial cells of Cajal, epithelium, and endothelium, and on our automated live-imaging pipeline for contraction frequency, amplitude, and coordination. We will integrate organ-level quantitative peristalsis measurements with high- content imaging and single-nucleus transcriptomics, yielding multimodal “fingerprints” that both reveal drug mechanisms and that will power predictive models for efficacy and GI-specific toxicity. We will pursue two aims: Aim 1—Engineer and rigorously validate peristaltic-HIOs for 96-well scalability, reproducibility, and cellular fidelity using design-of-experiments optimization and single-cell/spatial QC signatures. Aim 2—Establish a quantitative, screen-ready peristalsis assay and benchmark pharmacologic responsiveness to canonical neuromuscular modulators to define assay dynamic range and acceptance criteria. At the end of Phase I, we will have a validated human peristalsis model with locked manufacturing and QC, a reference pharmacology panel and assay performance metrics suitable for discovery, and an integrated “HIO atlas” and predictive models that accelerate hit identification, repurposing, and lead optimization.

NCATS - National Center for Advancing Translational Sciences

ABSTRACT A central barrier to next-generation immunotherapies is the scarcity of disease-specific extracellular targets. A major untapped source of targets arises from alternative splicing, which generates thousands of aberrant isoforms across many complex diseases. Many of these isoforms alter extracellular domains of transmembrane proteins, producing “splice surface antigens” (SSAs)—a novel class of epitopes with immense therapeutic potential. Despite strong biology rationale, no scalable, end-to-end platform exists to systematically discover, prioritize, and validate SSAs for drug development. This STTR Phase I will translate recent academic advances into the NeoSplice Discovery Engine™, a modular, disease-agnostic platform that 1) resolves full-length isoforms in heterogeneous samples, 2) scores candidates for surface exposure, safety, and tractability, and 3) confirms stable surface expression with targeted protein assays. Aim 1 will generate a ranked catalog of SSA candidates in a defined disease model, producing concise, partner-ready target dossiers. Aim 2 will establish a targeted validation workflow to verify isoform-specific surface expression across prioritized candidates and scale to larger panels. Phase I deliverables include a versioned scoring framework, prioritized SSA lists with per-target summaries, and validated assay panels suitable for therapeutic programs. This project lays the foundation to expand the actionable target space beyond canonical proteins. It will position us for Phase II lead generation, partnerships, and co-development of safer, more precise isoform-resolved therapies across multiple indications.

NCATS - National Center for Advancing Translational Sciences

SUMMARY Primary biliary cholangitis (PBC) is a chronic autoimmune liver disease in which the body’s immune system attacks the small bile ducts, leading to cholestasis (toxic bile accumulation), fibrosis, and ultimately liver failure. A hallmark of PBC is the activation of autoreactive B cells that produce anti-mitochondrial antibodies (AMAs), but the mechanisms that regulate B-cell activation within the liver microenvironment remain poorly defined. Recent work from our group and others indicates that liver lymphatic endothelial cells (LyECs), which line the lymphatic vessels running alongside bile ducts, play an active role in regulating immune cell trafficking and tolerance. In our preliminary analysis of single-cell RNA-seq data from human PBC livers, we identified striking upregulation of P2RY8, a G-protein–coupled receptor (GPCR), in both LyECs and B cells. P2RY8, an understudied gene listed in this RFA, couples to the Gα13–RhoA pathway to control immune cell positioning and is activated by S-geranylgeranyl-L-glutathione (GGG), a naturally occurring lipid metabolite in the mevalonate pathway. Although P2RY8 regulates B-cell migration in lymphoid tissues, its role in the liver or vascular endothelium has never been explored. This high-risk, high-reward R03 project will define how P2RY8 signaling reprograms LyEC phenotype (Aim 1) and how LyEC P2RY8 activity influences B-cell activation and AMA production (Aim 2). By uncovering a novel LyEC–B-cell communication axis, this work will establish a new conceptual framework for autoimmune cholangiopathies and lay the foundation for future studies exploring P2RY8 as a therapeutic target in PBC.

NCATS - National Center for Advancing Translational Sciences

Adenoid cystic carcinoma (ACC) of salivary glands is a rare, slow growing malignancy. Due to its insidious infiltrative growth pattern, ACC is often advanced by the time of clinical recognition, making it a difficult disease to treat. Current therapeutic options for primary localized ACC are restricted to surgery followed by radiotherapy with or without chemotherapy, as no systemic agent has been found to be effective in improving long-term disease control. Unfortunately, standard curative management has modest activity as most patients recur, largely due to ACC’s high tendency for perineural invasion and/or distant metastasis. Given its rarity, mechanisms that govern ACC pathogenesis remain poorly understood. Studies investigating new targeted therapies for ACC are hindered by the paucity of ACC specimens and preclinical cellular models. As a result, no FDA approved targeted treatments are currently available, and over 60% of the patients succumbing to the disease within 10 years of diagnosis. Development of the safe and effective targeted therapies is imperative for improving health outcomes and survival of patients with ACC. This proposal is specially designed in response to this notice of funding opportunity (NOFO) focusing on elucidating the role of understudied proteins in rare diseases. By coupling a large list of understudied proteins open for study under this NOFO with unique transcriptomic dataset of primary ACCs (a rare disease) and PandaOmics (a novel AI-driven therapeutic target discovery engine), we have selected HECTD2 and JMJD1C proteins as top-ranked putative therapeutic targets for ACC. While tangential previous findings pose HECTD2 and JMJD1C as potential drivers of certain types of cancer, their role in ACC tumorigenesis remains unknown, and thorough investigation is needed. In this project we will use a panel of authenticated and well characterized ACC cell lines and organoids, coupled with innovative proteomic approaches and comprehensive bioinformatics analysis of human specimens to address 3 objectives: (i) assess whether HECTD2 or JMJD1C deletion would induce anti-tumor activity in cellular models of ACC; (ii) map HECTD2 and JMJD1C protein interactome landscape; (iii) to define key HECTD2 or JMJD1C dependent signaling pathways that drive ACC tumorigenesis. These objectives are commensurate with the short-term nature of these research (one year), and specifically tailored to generate data critical for considering these proteins as potential candidates for therapeutic compound development against ACC. Given the devastating nature of ACC and dearth of effective treatment approaches, discovery of the previously unknown target candidates followed by the validation of their potential therapeutic activity in preclinical models (already available in our group) may ultimately translate to clinical trial settings and subsequently improve patients’ outcomes. As such, this pilot project is both innovative and clinically important.

NCATS - National Center for Advancing Translational Sciences

NabGen is developing an engineered and long half-life Protein M, a mycoplasma-derived protein, for blocking alloantibodies (inhibitors) for clinical applications and thus enabling the improved outcome of biological drug treatment in patients with antidrug antibodies (ADA), such as inhibitors from protein replacement treatment (PRT) in lysosomal storage diseases and hemophilia. The development of inhibitors to the infused proteins is the major complication in PRT. The costs of treatment for patients with inhibitors are 5-fold higher than patients without inhibitors. Additionally, patients with inhibitors suffer more severe diseases and hospitalization. Although many approaches have been used in clinical practice including deletion of B cells or T cells, high- dose of IVIG, induction of immune tolerance, these approaches often take long-time management and don’t work for each patient. Recently, we have demonstrated that Protein M binds to all immunoglobulin from any mammalian species with high affinity and is able to block immunoglobulin function. Our preliminary data shows that Protein M can reduce neutralizing antibody (Nab) activity by 100-fold in vitro and 1000-fold in animals when paired with a gene delivery vector, adeno-associated virus (AAV). Development of the Protein M-based strategy has significant advantages for blocking inhibitor/alloantibody activity, as Protein M: 1) has universal function, 2) has no effect on the biologics’ potency, 3) possesses high efficiency (100–1,000-fold prevention of inhibitors), 4) is capable of repeated administration, 5) with no severe side effects or complications seen in both small and large animals, and 6) potential low cost. Thus, to our knowledge, Protein M is the most effective approach to block inhibitors among all strategies studied so far. The wild-type protein M is not stable at room temperature. We have engineered one protein M mutant MG29 with increased thermo-stability and demonstrated that MG29 had a similar activity as wt protein M to block FVIII inhibitor activity in vitro and in vivo, in hemophilia A mice. To increase MG29 half-life, we have developed a fusion protein (MG64) with MG29 linked to an Fc domain and found the similar ability to block AAV Nabs when compared to MG29. This Phase I project seeks to explore the efficacy of fusion Protein M MG64 to protect FVIII treatment in hemophilia A mice with inhibitors. If successful, ultimately, this technology will extend the benefits to a greater population of patients with inhibitors and needing repeated administration of any biological therapy.

NCATS - National Center for Advancing Translational Sciences

This project focuses on defining the biochemical and cellular functions of B3GNT4 in support of the long-term goal of learning about its role in alpha-dystroglycanopathy and other human diseases. B3GNT4 is an understudied protein. It is highly expressed in the brain and the limited biochemical analysis that has been performed is consistent with it functioning as a Golgi-resident beta-1-3-N-acetylglucosaminyltransferase (beta- 1-3-GlcNAc-transferase) that participates in the biosynthesis of poly-N-acetyllactosamine (polyLacNAc) chains. A recent report described biallelic mutations in the B3GNT4 gene in a patient with progressive muscular weakening and brain atrophy but no mutations in genes known to be associated with muscular dystrophy. However, the mechanism by which B3GNT4 mutation could cause an alpha-dystroglycanopathy was not investigated. In this R03 project, we will conduct cell culture experiments and in vitro biochemical assays to obtain fundamental information about B3GNT4 in effort to elucidate possible mechanisms for its role in alpha- dystroglycanopathy phenotypes. We will establish neuronal and skeletal muscle cell lines with genetic knockout of B3GNT4, as well as cell lines with rescued overexpression of wild-type B3GNT4 or the disease-associated G160W mutant. We will also prepare purified, recombinant B3GNT4 catalytic domain as well as the corresponding G160W mutant. These reagents will be important tools for our aims and will be available to the research community. In the first aim, we will use these reagents to identify molecules glycosylated by B3GNT4. Specifically, we will determine whether alpha-dystroglycan is glycosylated by B3GNT4 and also identify any additional proteins glycosylated by B3GNT4. We will define the glycan structure that serves as a B3GNT4 acceptor as well the glycan structures that are produced by B3GNT4 activity. In the second aim, we define the impact of the G160W disease-associated mutation on B3GNT4. Using recombinant proteins, we will measure how G160W impacts B3GNT4 enzyme kinetics. Using cell culture experiments, we will evaluate how G160W affects B3GNT4 localization and binding partners. Studying the biochemical function of B3GNT4 will lead to new insights into (1) the neurological phenotypes of alpha-dystroglycanopathies and (2) the roles of B3GNT4 in other diseases.

NCATS - National Center for Advancing Translational Sciences

Project Summary/Abstract Many rare diseases result from single-gene mutations. Studying these disease-associated genes is crucial, as investigating their genotype-phenotype relationships can reveal the physiological roles of key cellular pathways relevant to both rare and common diseases. Because mutations in different genes within the same cellular pathways can produce similar pathological mechanisms, systematically investigating disease-associated genes within shared pathways and developing broadly applicable approaches to study them can greatly advance understanding of key cellular pathways and disease mechanisms. Responding to the funding opportunity for Pilot Projects Investigating Understudied Proteins Associated with Rare Diseases, this project aims to test the hypothesis that the understudied protein SCYL2 participates in clathrin-mediated endocytosis (CME) and that its loss of function disrupts endocytosis and neuronal differentiation, contributing to Arthrogryposis Multiplex Congenita type 4 (AMC4) pathology. This project serves as a starting point for a long-term objective: establishing a systematic framework for investigating the dynamics and functions of disease-associated endocytic proteins during development. AMC4 is a rare developmental disorder caused by loss-of-function variants in SCYL2, a putative kinase implicated in clathrin-mediated vesicle trafficking. AMC4 is characterized by multiple joint contractures and other neurogenic phenotypes, including brain malformations, agenesis of the corpus callosum, hypotonia, and epilepsy. Although SCYL2 interacts with coat proteins such as clathrin and AP2 adaptors, its role in CME remains unclear, partly due to limitations in approaches used to study endocytosis in prior decades. Understanding SCYL2’s function in endocytosis and neuronal development is essential for elucidating the cellular and molecular mechanisms underlying both CME and AMC4. Using human induced pluripotent stem cells (hiPSCs), genome editing, advanced live-cell imaging, and computational analysis, this study will apply newly developed quantitative pipelines that enable high-throughput analysis of endocytic protein dynamics as well as sensitive detection of CME defects through the following aims: Aim 1: Determine whether endogenous SCYL2 is recruited to endocytic sites. Aim 2: Test whether AMC4-associated SCYL2 variants affect endocytosis. Aim 3: Assess whether AMC4-associated SCYL2 variants impact neuronal endocytosis and differentiation. This project will generate genome-edited hiPSC lines as essential research tools and produce preliminary data elucidating the molecular mechanisms of SCYL2 in CME and neuronal development. The findings will advance understanding of CME regulation and AMC4 pathogenesis and establish broadly applicable methods for studying other disease-associated endocytic proteins in future research.

NCATS - National Center for Advancing Translational Sciences

Project Summary CACNA2D4 is the gene encoding a2d-4, an extracellular protein that is expressed primarily in the retina. Variants in CACNA2D4 are linked to a spectrum of inherited retinal disorders (IRDs) including cone dystrophies and retinitis pigmentosa. Despite clear evidence from human genetics and studies of knockout animals that a2d-4 is required for vision, it remains an understudied protein. The lack of tools to pinpoint the subcellular localization and modify the function of a2d-4 represents a major hurdle to understanding its role in IRDs. The goal of the proposed research is to develop antibody-based strategies to investigate the cellular functions of a2d-4 in normal and diseased states of the retina. We will use innovative approaches to generate renewable monoclonal antibodies and nanobodies that specifically recognize a2d-4 for use in electron and super-resolution light microscopy or to disrupt the interactions of a2d-4 with specific protein partners. The expected outcomes of the proposed research will contribute to the Pharos mission by shedding light on the localization and function of an understudied ion channel protein. The broader impact of this research is the discovery of mechanisms underlying a2d-4 function that could be targeted in novel disease-modifying therapies.

NCATS - National Center for Advancing Translational Sciences

Abstract DelAQUA Pharmaceuticals is developing a non-PEG, stealth, hyper-loaded polymeric micelle (PM) approach for drug coadministration with P-glycoprotein (Pgp) inhibitors to address the barriers of poor solubility and low oral bioavailability due to Pgp efflux. Oral administration is the preferred drug delivery route based on convenience, safety, cost-effectiveness, improved access to care, and high patient compliance rates. However, physiological and biochemical challenges exist for effective oral drug administration. Poor aqueous solubility significantly contributes to low bioavailability, impacting about 40% of approved drugs and upwards of 90% of newly synthesized drug candidates in development. Oral bioavailability is also often limited by Pgp-mediated efflux in the gut. It significantly limits the oral bioavailability/uptake of many lipophilic drugs, mediates their direct excretion into the intestinal lumen, and results in drug resistance. This absorption barrier is estimated to impact >25% of lipophilic drugs, which comprise most small-molecule drugs in pre-clinical development. Paclitaxel (PTX), an important drug in treating many cancers, suffers from poor solubility and Pgp efflux. The PTX standard of care, Abraxane®, only addresses the solubility problem using an albumin-based nanoparticle, still requires intravenous administration, and the response rate remained relatively low (33%) in metastatic breast cancer patients. As such, an unmet need remains for innovative solutions for oral delivery of PTX and other Pgp substrate drugs with poor solubility. DelAQUA’s platform technology utilizes water-soluble and nontoxic polymers arranged in an amphiphilic triblock copolymer structure to incorporate poorly soluble drugs into biocompatible, chemically versatile, and tunable PMs. The molecular interactions between the poly(2-oxazoline) (POx) hydrophilic and hydrophobic repeat units of block copolymers drive the formation of a hydrophilic corona with a hydrophobic core to support improved solubilization and superior loading. The POx platform has been used to package a variety of drug combinations and has co-delivered two drugs in a single micelle with increased tumor distribution of both drugs. This Phase I project seeks to establish the feasibility of the PM platform for co- formulating poorly soluble Pgp substrate drugs like PTX with Pgp inhibitors like elacridar to enhance oral bioavailability through the following Specific Aims: 1) develop an optimized oral micelle formulation for the co- administration of PTX and Pgp inhibitors, 2) assess the oral bioavailability and tumor accumulation of PTX with Pgp inhibitor micelle formulation in tumor-bearing mice, and 3) validate the therapeutic efficacy of an oral micelle PTX:elacridar formulation in a mouse model of breast cancer. This work will equip DelAQUA with a well- characterized PM product for oral delivery of PTX with superior bioavailability and inform future work optimizing the formulation and expanding the platform for the solubilization of other Pgp substrate active pharmaceutical ingredients (APIs) for much-needed orally delivered therapeutics, yielding improved treatment outcomes and improved adherence and access to care.