Andrew Nguyen - US grants

Project Summary Progranulin is a lysosomal and secreted protein that contains multiple cysteine-rich granulin domains; its precise molecular function remains unknown. Progranulin (GRN) mutations are causal for frontotemporal dementia (FTD), which is a devastating disease with a mean survival of 3.8 years from diagnosis and no cure currently available. Since progranulin-deficient FTD is a disease of haploinsufficiency, strategies aimed at increasing progranulin levels are feasible therapeutic approaches. We recently generated a knock-in mouse model of FTD harboring the common patient nonsense mutation GRNR493X, and we established that the nonsense-mediated mRNA decay (NMD) pathway contributes to the markedly reduced mutant progranulin mRNA levels in this mouse model, as wel`l as in patient-derived fibroblasts containing the GRNR493X mutation. Moreover, our cell-based studies indicate that the progranulin R493X mutant protein is functional when re- expressed in progranulin-deficient cells. Based on these results, we hypothesize that inhibiting NMD-mediated degradation of the mutant GrnR493X mRNA would increase functional progranulin levels and thereby improve FTD-associated behavioral changes and neuropathology. We will test this hypothesis in the following two specific aims: Aim 1) Test in vivo efficacy of ASOs that block NMD-mediated degradation in the GrnR493X knock-in mouse model of FTD, and Aim 2) Design and test ASOs that target NMD-mediated degradation of the human GRNR493X mRNA. Completion of these studies will generate important preclinical data that provide insights into the utility of ASO-based NMD inhibition as a therapeutic approach for progranulin-deficient FTD.

An award is being made to The City University of New York (CUNY), Queensborough Community College (QCC) campus (Department of Biological Sciences & Geology), to purchase a LI-COR Odyssey Imaging System. This multitasking imaging system allows for the visualization of Quantitative Western blots (WB), In-Cell Western Assays, Viral Titration Assays, and Nucleic Acid Gel Imaging – to name a few of its applications. QCC is one of the seven community colleges within CUNY and serves a diverse population of first-generation college students and admits many students who would otherwise not have access to higher education. 84% of QCC students are African American, Asian, or Latino, and more than 50% come from families with incomes of less than $25,000. More than a third of the Biological Sciences and Geology department’s faculty members mentor students in research techniques and ethics every semester. Our faculty members are exceedingly invested in the success of our students as they embark on their careers in STEM fields. Acquisition of this system will complement the campus’s undergraduate research core facility, which in part includes a real-time PCR equipment, a FACS machine, and a confocal microscope. All instrumentation in the core facility allows faculty to introduce students to cutting-edge experimental methodologies and helps prepare them for entry into the STEM workforce. The newly acquired LI-COR Odyssey instrument will also be an integral part of the educational training in the Biotechnology Program and the newly formed Associates Program in Biology at QCC, thus impacting even more students.

The LI-COR Odyssey is an essential part of the continuous research effort and teaching curriculum at QCC and its sister campus, Queens College. Its acquisition will support a variety of diverse projects. These include studies of (a) the molecular regulation of STAT3 in cell-cell interactions, (b) regulation of host amino acid transporters by an intracellular parasite, (c) examination of conserved root development genes in Selaginella (d) inhibition of DnaK by intrinsically unfolded polypeptide substrates in Escherichia coli, (e) investigation of cancer cell gene expression after treatment with single-walled nanotubes, (f) the molecular regulation of Cnidarian behaviors, (g) investigating novel proteins in obesity and associated comorbidities, (h) role of CDX1 as a regulator of colon cancer sensitivity to chemotherapeutic drugs and (i) cardiovascular health-related research. Data generated using the instrument will be disseminated by faculty and students through peer-reviewed journal publications and at both regionally and nationally held professional and scientific conferences. Procuring this system will thus open new frontiers of research for several of QCC’s most recently recruited early career investigators and their students.

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.