Spatial Characterization of Mechanosensory and Chemosensory Receptor Expression in Drosophila’s Digestive tract

Obesity constitutes one of the greatest public health challenges, having a high prevalence and increasing the risk of serious complications. Obesity is essentially caused by an imbalance of energy intake and expenditure, which is under the tight regulation of the nervous system. The goal of this project is to investigate the genetic causes of abnormal food intake and eating regulation, by studying the different mechanosensory and chemosensory receptors present in the fruit fly Drosophila melanogaster’s digestive system. Tiffany will perform immunofluorescent staining and imaging using confocal fluorescence microscopy to identify the specific neurons that are involved in gut-brain communication. Following identification, Tiffany will analyze the spatial distribution of neural innervation and characterize the physiological and behavioral functions of specific neurons. The outcomes will further the understanding of neuron function and feeding regulation mechanisms.

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Genetic Modulation of Two Phosphatases, Ptpn6 and Ptpn22, to Examine their Role in NK Cell Desensitization

Natural killer (NK) cells play an important role in the innate immune system in eliminating viruses and tumors. Unfortunately in environments lacking class I major histocompatibility complex (MHC-I) proteins, as is the case in 40-90% of human tumors, the NK cells can become desensitized and unresponsive. In her project, Seungwon will modify the expression of two phosphatases, Ptpn6 and Ptpn 22, that have been found to be expressed at higher levels in anergic NK cells to study the molecular mechanisms of NK cell desensitization. Findings have the potential to guide NK cell cellular therapy.

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The Role of Mannan Associated Serine Protease (MASP-1) in Immune Recognition of Human Cytomegalovirus (HCMV)

Human Cytomegalovirus (HCMV) infects 60-90% of humans globally, and while infections are largely asymptomatic, they can be severe or fatal in immunocompromised persons. No vaccines exist to prevent HCMV infection due to an incomplete understanding of the viral mechanisms used to evade host immunity and establish lifelong persistence. Complement is a system of proteins present in blood that serves as an initial line of defense against microorganisms; however, little is known about the role of complement in protection against HCMV. For this project, Hector will investigate the role of MASP1 (a complement activating protein) in HCMV immunity using a yeast-two-hybrid approach to screen for protein-protein interactions against an HCMV gene library. Identification of novel HCMV-complement interactions has the potential to inform the development of vaccines and novel drugs.

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The Role of ORF69 in Lytic Reactivation of Kaposi’s Sarcoma-Associated Herpesvirus

Kaposi’s sarcoma-associated herpesvirus (KSHV) establishes lifelong infections and can cause various cancers in immunosuppressed individuals. KSHV lies dormant in infected individuals’ cells and reactivates intermittently to cause disease and promote transmission. Current scientific literature lacks an understanding of the KSHV gene open reading frame 69 (ORF69). In this project, Christian will employ a two-step mutagenesis technique in E. coli to generate ORF69 knockout and revertant mutants in a KSHV bacterial artificial chromosome (BAC). After transfecting a Kaposi’s sarcoma cell line with the BAC vectors, inducing lytic reactivation, and infecting 293T cells with supernatant from the induced cells, Christian will be able to determine if ORF69 is essential in the KSHV life cycle. The results have the potential to inform the development of novel drugs and immunotherapies for KSHV transmission and pathogenesis.

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Identifying Host Factors Critical for Dengue Virus NS1 Internalization and Pathogenesis

Dengue disease is caused by four serotypes of dengue virus (DENV), and associated symptoms can range from undifferentiated fever to severe vascular leakage. DENV nonstructural protein 1 (NS1) was recently found by the Harris laboratory and others to be a key factor in causing the endothelial barrier dysfunction that leads to vascular leakage, but the mechanism is not yet completely understood. Richard will focus on identifying possible host factors critical for NS1-induced pathology by generating a list of host factors of interest from pilot gene expression analyses, and then validating their role in endothelial dysfunction using a murine dermal leak model. Richard will then assess the therapeutic potential of small molecule inhibitors of host factors critical for NS1-mediated pathology and create modified cell lines to better understand the overall mechanism.

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