Grants

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY AND ABSTRACT Cytomegalovirus (CMV) is a common herpesvirus that establishes latency for the lifetime of the host and drives immune dysregulation. The lung is an important reservoir for CMV in both humans and mice. CMV infection can reactivate during episodes of immunosuppression, with age, and from direct tissue injury. Importantly, CMV reactivation is associated with increased risk for death following lung transplantation, acute respiratory distress syndrome (ARDS) and SARS-CoV-2 pulmonary disease. Natural killer (NK) cells are effector innate immune cells that can recognize damaged or missing-self. They play a central role in mitigating CMV infection with long- lived tissue-resident, CMV-specific and adaptive abilities. Our prior publications have established novel roles for NK cells in mediating acute lung injury and found increased prevalence of CMV-specific NK cells in human lungs preceding injury. Our preliminary data identify tissue-resident NK cell populations as key effectors of airway- centric acute lung injury. Moreover, we found that adaptive-like NK cells are increased in tissue surrounding chronically-injured lung transplant airways. Identifying whether adaptive NK cell differentiation occurs locally in the lung in response to CMV reactivation and drives lung damage represents a fundamental gap in knowledge. Based on our observations, we hypothesize that local CMV-specific NK cell immune responses potentiate lung damage after transplant. Across 2 independent Aims we will use mouse models of CMV reactivation and observations in 2 established human lung transplant cohorts to investigate mechanisms of adaptive NK cell lung injury. Aim 1 will determine the impact of CMV reactivation on the adaptive NK cell response in the mouse and human lung. We will use 2 lung transplant mouse models and a previously recruited cohort of 56 lung transplant recipients’ bronchoalveolar lavage and tissue samples to measure NK cell phenotypes and spatial distributions relative to CMV activity and to probe the mechanism whereby lung adaptive NK cells develop in the allograft. In a non-interdependent fashion, Aim 2 will determine the mechanisms through which adaptive NK cells potentiate acute injury and their role in establishing allograft tolerance using established orthotopic allogeneic mouse lung transplant models. These proposed innovative approaches will examine the novel concept that tissue injury potentiates geographic residency of CMV-specific adaptive NK cell populations, raising the allograft immune tone and the risk for lung allograft damage. Work resulting from this proposal will be broadly applicable to other solid organ transplant recipients and patients suffering from acute respiratory distress syndrome and acute exacerbations of chronic lung disease. Importantly, we will identify: mechanisms of novel biology, cell populations that may risk stratify clinical outcomes, and new rational targets for future prevention and treatment of lung allograft syndromes.

Up to $795K

2031-03-31

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADAR1 promotes Okazaki fragment maturation and is a target for breast cancer therapy

NCI - National Cancer Institute

ABSTRACT To initiate lagging strand DNA synthesis, the polymerase α-RNA primase complex synthesizes RNA primers, which are composed largely of adenine (A) and cytosine (C) residues that start each of the millions of Okazaki fragments required for eukaryotic chromosome replication. RNA primers are synthesized in an error prone manner. Adenosine-to-inosine (A-to-I) modifications are common RNA editing events performed by adenosine deaminase acting on RNA (ADAR1). A consequence of A-to-I editing is that I is interpreted as guanine (G), functional A-to-G substitutions and correction of A:C mismatches within pre-mRNAs, mRNAs, and noncoding RNAs. There are two ADAR1 isoforms, ADAR1p150 and ADAR1p110, which are predominantly localized to the cytoplasm and nucleus, respectively. By modifying cytoplasmic double-stranded viral RNA, ADAR1 p150 suppresses interferon signaling. The functions of nuclear ADAR1p110, however, have been understudied. We have found a novel role for ADAR1p110 in editing of RNA primers during DNA replication. ADAR1p110 is overexpressed in breast cancer, particularly triple negative breast cancer (TNBC), an aggressive subset characterized by elevated levels of replication stress. Our preliminary data suggests that ADAR1p110 localizes with primase at nascent DNA, and that ADAR1p110 A to I editing of nascent DNA suppresses DNA gap formation and stabilizes the replication fork. Consistently, loss of endonucleases FEN1 and DNA2, key enzymes in Okazaki fragment maturation, were synthetic lethal with ADAR1p110. Poly (ADP-ribose) polymerase (PARP) inhibitors induce the accumulation of excessive single stranded DNA gaps in the lagging strand of replicating DNA in cancer cells, resulting in cancer cell death. PARP inhibition is approved for BRCA1 and BRCA2 associated breast cancers. However, many patients have inherent or acquired resistance, and PARP inhibition is not an option for patients without BRCA1 or BRCA2 alterations. We found that genetic deletion of ADAR1 p110 or inhibition of ADAR1p110 dimerization with a novel ADAR1p110 specific small molecule inhibitor identified through virtual screening resulted in DNA gaps and tumor specific hypersensitivity to PARP inhibition and other replication gap targeted therapy. Further, we have found that the multifunctional protein, APEX1, may regulate this new ADAR1p110 role. We hypothesize that ADAR1p110 stabilizes replication forks by editing mis- matched RNA:DNA hybrids and suppressing replication gaps and is a target to overcome resistance to breast cancer therapy. Aim 1 will investigate ADAR1p110 RNA primer editing and Okazaki fragment processing. Aim 2 will examine the molecular function of APEX1 in ADAR1p110 regulation. Aim 3 will evaluate ADAR1p110 as a target for therapeutically resistant breast cancer.

Up to $590K

2031-04-30

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADB Asia Pacific Project Preparation Facility (AP3F)

Asian Development Bank

Supports governments in preparing bankable infrastructure projects through feasibility studies, transaction advisory, and capacity building for PPP projects.

$250K – $1.5M

infrastructureproject preparationpublic-private partnerships

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADB Clean Energy Financing Partnership Facility (CEFPF)

Asian Development Bank

Supports clean energy investments in ADB developing member countries through grants for project preparation, capacity building, and policy reform.

$200K – $3M

clean energyrenewable energyclimate change+1

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADB Financial Sector Development Partnership Fund

Asian Development Bank

Supports financial sector development in ADB developing member countries through technical assistance grants for financial inclusion, capital market development, and regulatory reform.

$200K – $2M

financial inclusioncapital marketsregulatory reform

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADB Japan Fund for Poverty Reduction (JFPR)

Asian Development Bank

Funded by the Government of Japan, provides grants for innovative poverty reduction and social development projects in ADB developing member countries.

$500K – $3M

poverty reductionsocial developmentcommunity development

Free to search & build · $99 one-time to unlock the application pack · No subscription

ADB Urban Climate Change Resilience Trust Fund (UCCRTF)

Asian Development Bank

Helps cities in Asia build climate resilience through integrated urban planning, infrastructure investment, and community-based adaptation.

$500K – $5M

climate resilienceurban developmentinfrastructure+1

Free to search & build · $99 one-time to unlock the application pack · No subscription

Addiction Application Gradebook: Fostering greater public health impact from mHealth applications for SUD

NIDA - National Institute on Drug Abuse

Project Summary Recent estimates show 85% of United States adults own a smartphone and 56.6% report using a mobile health application in the past year. Mobile health applications targeting substance use and use disorder can fill numerous treatment gaps, such as providing broader access, enhanced flexibility, and opportunity for individual self-determination. At the same time, app quality is variable, poorly understood, and both academic and commercial developers are flooding the marketplace. The proposed Phase One STTR project will establish foundation research and development for the Addiction Application Gradebook, a web platform dedicated to the description, evaluation, and market assessment of smartphone- and web-based applications for substance use disorder (SUD) information/prevention, harm reduction, treatment, and recovery. While increased availability of mobile apps for SUD is an important step forward, rapid growth coupled with the absence of an infrastructure for organizing, evaluating, and tracking these products has resulted in a large and difficult to navigate landscape. Therefore, the Phase One aims of this proposal are to conduct research and development in three key areas related to SUD app 1) Identification, 2) Evaluation, and 3) Tracking to yield a minimum viable product design that would set the stage for Phase Two development, testing, and commercialization. The proposed project outlines clear and measurable milestones for success that include a procedure for ongoing app identification, assembling an expert Delphi panel, and creating a novel SUD app evaluation framework as well as a model for understanding the SUD app innovation pipeline. The culmination of project milestones will be foundation data for a web platform with functionality for no cost public education (i.e., the public can search and find the right app for a given SUD need) and commercialization via a business-to-business membership model (i.e., app developers, employee assistance contractors, mHealth investors and scientific funders can obtain market assessments, product comparisons, and innovation trajectory analytics). The proposed project is well aligned with NIDA Strategic Priorities, particularly Areas 2 and 5 related to prevention, treatment, harm reduction, recovery, and scientific innovation translation. The Addiction Application Gradebook can fill several needs in a rapidly growing mHealth market valued at 93 billion dollars in 2023. There is currently no comparable product while there is commercial potential for the general business model we propose (i.e., app selection, market research, and analytics). Phase One is focused on building the platform scientific base; in Phase Two we will prioritize web-design, incorporation of artificial intelligence capabilities, and commercialization.

Up to $314K

2027-05-31

Free to search & build · $99 one-time to unlock the application pack · No subscription

Adding spatial resolution and technical improvements to a novel single-cell DNA methylation sequencing technology

NHGRI - National Human Genome Research Institute

ABSTRACT DNA methylation (DNAme) is a core layer of epigenetic regulation with central roles in the establishment and maintenance of cellular identity. Methods to profile DNAme at single-cell resolution are needed to elucidate epigenetic networks governing cell state in healthy tissues and understand their dysregulation in disease and aging. However, existing methods for single-cell methylome profiling are highly inaccessible, and no methods to experimentally profile both DNAme and spatial location currently exist. I have developed a novel method for single-cell DNAme profiling that leverages the widely available 10x Single Cell Multiome and NEB EM-seq kits, which I have named droplet-implemented single-cell DNA methylation sequencing (discDNAme). Applied to nuclei isolated from mouse brain, discDNAme recovered ~3,000 high-quality methylomes that clustered into clearly separated neuronal and non-neuronal subtypes and displayed stereotyped patterns of CpG dinucleotide methylation around key genomic features. However, these measurements lack spatial information on the native tissue contexts of profiled cells, and the protocol’s per-cell library size is lower than current gold-standard methods for measuring single-cell DNAme. I propose (1) development of a spatially resolved single-cell DNA methylation technology by combining discDNAme with slide-tags, a platform for spatially resolved single-nucleus RNA- and/or ATAC-seq developed by our group. To integrate slide-tags with discDNAme, I will develop a protocol in which the “spatial barcode oligos” we use to position nuclei are physically separated from genomic DNA prior to unmethylated cytosine conversion, benchmark this technology in the mouse hippocampus, and apply it to study glioblastoma multiforme. I further propose (2) experiments to improve and benchmark my discDNAme technology. I will systematically test various independent approaches to increase library complexity at different steps of the discDNAme protocol, combine these optimizations into a second-generation protocol, and benchmark this against our original protocol and other leading methods for single-cell DNAme profiling. Completion of this proposal will result in (1) the first high-resolution method to measure spatially resolved single- cell methylomes—a major advancement in spatial omics technologies—and (2) an accessible yet capable tool for single-cell DNAme profiling that will open single-cell DNAme studies to the broader single-cell community.

Up to $44K

2028-01-31