Pilot Projects

Current Pilot Projects

The following pilot projects are currently being funded for the 2017 grant year:

Principal Investigator:  Dr. Craig Vander Kooi

Department of Molecular and Cellular Biochemistry

Title:  Function of GIPC3 in Hearing Loss

Abstract:  The GIPC family of proteins function as key molecular scaffolds that are critical in the physical integration of multiple steps of signaling from receptor activation to intracellular signaling. GIPC3 plays an essential role in the auditory system, with mutations in GIPC3 causing inherited nonsyndromic hearing loss. Despite this unique and crucial function in the auditory system, the molecular function and why mutations in the GIPC3 gene leads to hearing loss, is unknown. We will define the structural basis for GIPC3 function, and the mechanism(s) by which the eleven reported patient mutations lead to inherited hearing loss. Defining the structure and functional interactions of GIPC3, and the molecular basis for dysfunction, requires an integrated structural, biochemical, and physiological approach that form the heart of our proposed work. In Aim 1, we will determine the basis for GIPC3 coupling to the NMDA glutamate receptor. Aim 2 will focus on conformational dynamics associated with activation of GIPC3, and how this leads to engagement of the Myo6 motor protein. Taken together, these studies contribute to our long-term goal of understanding the physical mechanisms underlying the role GIPC3 in normal physiological function, and will inform ongoing efforts to understand and ameliorate GIPC3 dysfunction in disease.

Principal Investigator:  Dr. Adam Bachstetter

Spinal Cord & Brain Injury Research Center

Title:  Role of MK2 Following Traumatic Brain Injury

Abstract:  This 2-year pilot award will generate preliminary data investigating the role of MAPK-activated protein kinase 2 (MK2) following traumatic brain injury (TBI). Despite multiple attempts, no drug has earned FDA approval for TBI. Control of neuroinflammation is a promising avenue to achieve neuroprotection and improve patient outcomes. Still, there is an urgent need to identify the molecular mechanisms that regulate inflammation, so selective therapeutic agents can be developed. Our prior work has demonstrated that the mitogen-activated protein kinases (MAPK) pathway is involved in detrimental neuroinflammation following injury or in neurodegenerative disease [1-4]. From our prior work we have recently identified the protein MK2, which we postulate maybe an enzyme specific to the detrimental proinflammatory response to TBI. Using a mildly closed head injury model in mice we will address our overall hypothesis: MK2 deficiency will prevent TBI induced neurological deficits. If successful, we will have identified a novel therapeutic target that regulates neuroinflammation following TBI, which will lay the foundation to garner NIH funding to explore the pathway in greater detail.

Principal Investigator:  Dr. Jessica Blackburn

Department of Molecular and Cellular Biochemistry

Title:  Single Cell Characterization of Leukemia Stem Cells

Abstract:  Patients with relapsed Acute Lymphoblastic Leukemia (ALL) have a poor prognosis because their therapeutic options are limited. Relapse is thought to arise from
leukemia stem cells, which have the unique ability to self-renew to reform tumor from a single cell. These cells are rare in human disease and mouse models, making up <0.0001% of the total leukemic cell population. Because we do not yet know how to reliably isolate these cells from the bulk of the tumor, leukemia stem cells are not well characterized, despite their importance in ALL progression and relapse. We have developed a zebrafish model of ALL in which leukemia stem cells make up ~10% of the tumor cell population. The goals of this proposal are to characterize leukemia stem cells from T-ALL functionally through single-cell transplantation and genetically through single-cell RNA sequencing. With this preliminary data, we will apply for R01 funding from NCI to scale up single cell sequencing efforts, and to use zebrafish ALL models for in vivo drug screens to identify compounds that target leukemia stem cells. These data will ultimately provide new and important insights into cancer biology and will lead to the identification of new targets for anti-cancer therapies.

Principal Investigator:  Dr. Konstantin Korotkov

Department of Molecular and Cellular Biochemistry

Title:  Structural and Biochemical Perspective of Cell-wall Biosynthesis in Gram-positive Bacteria

Abstract:  Because of the growing prevalence of antibiotic-resistant bacteria there is an urgent need for new antibiotics and novel approaches for treating infections. Cell wall biosynthetic machinery and cell wall associated carbohydrates are preferred targets for the discovery of novel antimicrobials. Group-specific carbohydrates are a major cell wall component of pathogenic streptococci. They play a critical role in bacterial survival and they have important applications as diagnostic tools, vaccines, and drug targets. The goals of this project are to understand the biosynthesis mechanism of group-specific carbohydrates expressed by human pathogen Streptococcus pyogenes (Fig. 1). Our preliminary and published findings reveal insights into S. pyogenes carbohydrate composition and biosynthesis and have developed unique bacterial strains, molecular tools and novel non-destructive methods of carbohydrate extraction and characterization. To accomplish our goals, we propose to utilize the methods of structural biology, streptococci genetics, and polysaccharide and lipid biochemistry. Successful outcomes will guide studies of biogenesis of Group-specific carbohydrates expressed by other important Gram-positive bacterial pathogens and development of novel strategies to treat Grampositive bacterial infections.

Principal Investigator:  Dr. Trevor Creamer

Department of Molecular and Cellular Biochemistry

Title:  Interplay Between Calcineurin, Rcan1, and Tau in Alzheimer's Disease

Abstract:  Alzheimer’s disease (AD) is estimated to affect one in nine individuals in the United States. AD is characterized by deposition of proteinaceous plaques and neurofibrillary tangles in affected brain tissue, the effects of which lead to cognitive decline and ultimately death. Tangles arise from the aggregation of hyperphosphorylated forms of the microtubule-binding protein tau. The origins of tau hyperphosphorylation are currently unknown but have been hypothesized to involve decreases in phosphatase activity, including that of the Ser/Thr phosphatase calcineurin (CaN). CaN is of note because it acts upon residues (T231 and S262) known to be important for regulation of binding to microtubules, as well as priming tau for additional phosphorylation. Our overall hypothesis is that CaN: tau interactions are disrupted in the AD brain as a result of increased expression of regulator of calcineurin 1 (Rcan1), an endogenous protein inhibitor of CaN. The specific objectives described in this proposal are to determine the molecular basis of Rcan1’s inhibition of CaN activity and determine the regions within tau responsible for binding to CaN. The results of these studies will be used as preliminary data in future grant proposals on the role CaN plays in the regulation of the microtubule-binding properties of tau and how overexpression of Rcan 1 contributes to the formation of neurofibrillary tangles.