Benjamin R. tenOever - US grants

DESCRIPTION (provided by applicant): Lowering intraocular pressure (IOP) slows or stops the loss of vision in primary open-angle glaucoma (POAG) patients, including in those with normal IOP. Unfortunately, current medical treatments are usually inadequate and do not target the conventional outflow pathway; the main route of aqueous humor egress and the site of extra resistance in POAG patients with elevated IOP. Presently in clinical trials are two new classes of drugs that inhibit the contractile machinery of conventional outflow pathway cells, effectively lowering IOP. Although promising, such drugs have limitations: they are disruptive, non-specific, do not restore tissue function, and are dependent on patient compliance for daily administration. An attractive therapeutic alternative for POAG are molecular therapeutics such as microRNAs (miRNAs) that achieve prolonged modulation of various biological functions such as cell contractility because of their ability to regulate entire networks of genes. Our group has recently identified miR-200c as a strong candidate to control IOP based on its ability to inhibit contractio of trabecular meshwork cells and lower IOP in vivo. To improve delivery of miRNAs in vivo, we have developed a new class of viral vectors, including influenza-associated-virus viral-like-particles (IAV-VLP) that overcome the limitation for effective in vivo delivery of miRNAs that result from overloading of the miRNA biogenesis components. Thus, we hypothesize that miR-200c regulates conventional outflow via modification of contractile state of trabecular and/or Schlemm's canal cells. We also hypothesize that over expression of miR-200c using IAV- VLP decreases IOP in normotensive animals and prevents the pathologic increase in IOP in experimental models of glaucoma. To test our hypothesis we have constructed three specific aims designed to (i) Assess the role of miR-200c on the regulation of cellular contractility of HTM and SC cells; (ii) Examine the effects of miR-200c on aqueous humor outflow function and IOP in perfused human anterior segments and living rat eyes; (iii) Test whether increased expression of miR-200c in the cells of the outflow pathway can prevent or restore the increase in IOP in experimental models of glaucoma. Taken together, results obtained from these investigations will establish feasibility of miRNA therapy for ocular hypertension in glaucoma, optimize delivery of miRNAs to conventional outflow tissues, identify new genes regulated by miRNA-200c responsible for IOP lowering and generate novel animal models for studying glaucoma.

Abstract. Recent clinical data has suggested a bidirectional interaction between Coronavirus disease 19 (COVID-19) and diabetes. Individuals with diabetes and severe obesity are more likely to be complications, and have a higher COVID-19 mortality rate symptomatic . Conversely, new-onset diabetes and severe , are at a higher risk for metabolic complications of pre-existing diabetes have been observed in COVID-19 patients. Thus, there is a strong need to understand the pathology and mechanism of pancreatic dysfunction in COVID-19 patients. Here, we demonstrate the detection of SARS-CoV-2 in pancreatic endocrine cells in autopsy samples from COVID-19 patients. Single cell RNA-seq and immunostaining confirmed that multiple types of pancreatic islet cells are susceptible to SARS-CoV-2, eliciting a cellular stress response and the induction of chemokines. Upon SARS- CoV-2 infection, beta cells show a decreased expression of insulin and the increased expression of alpha and acinar cell markers, including glucagon and PRSS1/trypsin1, respectively, suggesting cellular transdifferentiation. Hyperion technology to examine the pathogenesis of autopsy samples of COVID-19 patients. In addition, we will use human islets and a vascularized human pancreatic organoid models to systematically evaluate the role of direct infection and paracrine inflammation signal on human endocrine cells cellular identities, function and survival. In the proposal, we will apply state-of-art Through this study, we would expect to provide a systematic overview of the pathological changes in the pancreas of COVID-19 patients, as well as a detailed mechanism to understand endocrine cell dysfunction, which will pave the road to the development of novel therapy to protect endocrine cell function in COVID-19 patients.