2015 — 2020 |
Yadav, Smita |
K99Activity Code Description: To support the initial phase of a Career/Research Transition award program that provides 1-2 years of mentored support for highly motivated, advanced postdoctoral research scientists. R00Activity Code Description: To support the second phase of a Career/Research Transition award program that provides 1 -3 years of independent research support (R00) contingent on securing an independent research position. Award recipients will be expected to compete successfully for independent R01 support from the NIH during the R00 research transition award period. |
Role of Autism Susceptibility Gene, Taok2 Kinase, and Its Novel Substrates in Synaptogenesis @ University of California, San Francisco
? DESCRIPTION (provided by applicant): Dendritic spines are specialized actin-rich protrusions that serve as primary recipients of most excitatory synapses in the brain. Spines are extremely dynamic, exhibiting diverse structural and functional changes during development, in response to stimuli, as well as in learning and memory. While there is substantial evidence that several neurodevelopmental and psychiatric diseases converge on a common theme of aberrant spine formation, the mechanisms of spine formation and how its dysfunction relates to disease is unclear. TAOK2 is a serine/threonine kinase implicated in neuronal development, and is one of the genes present in the 16p11.2 genomic locus. Deletion of this region is the most common risk factor associated with autism spectrum disorder (ASD). Despite its relevance in neuronal development and to ASD, the physiological neuronal substrates of TAOK2 kinase are not known. It is unclear how TAOK2 signaling mediates spine development and how an imbalance in TAOK2 gene dosage might contribute to neuronal and behavioral alterations associated with ASD. Using a combination of innovative approaches, this proposal aims to delineate the mechanistic role of TAOK2 kinase during synaptogenesis and to understand how dysfunction in this signaling pathway might contribute to disease. As a postdoctoral fellow in Dr. Yuh-Nung Jan's laboratory at UCSF, I have gained skills in neurobiology, mass spectrometry, and chemical-genetics in addition to my graduate background in cell biology and biochemistry. For the experiments proposed in this application, I seek interdisciplinary mentored training in stem cell technology, super-resolution microscopy, and quantitative proteomics. I have assembled a team of mentors with expertise in: mass spectrometry and proteome-scale data acquisition and analysis (Dr. Al Burlingame), iPSC technology (Drs. Kathryn Ivey and Lauren Weiss), neurodevelopment and ASD (Dr. John Rubenstein), and super-resolution microscopy (Dr. Bo Huang). This training will allow me to achieve the following aims: 1) systematic investigation of TAOK2 localization during spine development using super-resolution STORM imaging, 2) identification and characterization of direct novel targets of TAOK2 kinase using a chemical-genetic approach, and 3) characterization of the TAOK2 pathway in ASD pathology using iPSC derived neurons from 16p11.2 deletion patients. With the new training and information acquired in the K99 phase, I will extend the scope of my research in the R00 phase. I will analyze the role of the identified TAOK2 phosphorylation targets during neuronal development. I will also perform phosphoproteomic analysis of 16p11.2 deletion iPSC-derived neurons to identify global changes caused by imbalances in TAOK2 dosage, which may reveal clinically relevant insights into the pathogenesis of ASD. The training period afforded by the K99/R00 award will greatly facilitate my long-term goal as an independent investigator to elucidate signaling pathways involved in dendritic spine formation during normal and disease states.
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1 |
2020 — 2021 |
Yadav, Smita |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Taok2 Kinase Signaling in Human Neural Stem Cell Development @ University of Washington
Increasing evidence indicates that aberrations in neural stem cell proliferation and early neurogenesis are critically involved in the pathogenesis of neurodevelopmental and psychiatric disorders. Despite being implicated in pathophysiology of several neurological diseases, the mechanisms through which human kinome controls neurogenesis and how its dysfunction manifests in disease remain major gaps in the field of neurodevelopmental biology. In this proposal, we will investigate the role of an autism susceptibility gene, TAOK2, which encodes a serine threonine kinase, in human neural stem cell development and function. Both loss-of-function and activating TAOK2 mutations have been associated with autism spectrum disorders (ASD). Further, TAOK2 is one of the genes in the 16p11.2 genomic locus, copy number variation (CNV) of which is the most prevalent genetic risk factor associated with ASD. While work by us and others has demonstrated compelling evidence that TAOK2 is important for neuronal and synaptic development, the role of TAOK2 in human neural progenitor cell (NPC) development and differentiation has not been investigated. Further, the contribution of TAOK2 in the pathology associated with 16p11.2 copy number variation is unknown. Based on the strength of our preliminary findings, the central hypothesis of this research proposal is that TAOK2 kinase orchestrates a signaling hub at the centrosome that regulates human neural progenitor stem cell development, and that perturbation of this signaling pathway contributes to pathogenesis of ASD. We will elucidate the role of TAOK2 in NPC development through use of 2D and 3D human induced pluripotent stem cell (hiPSC) models (Aim1). Next, mechanisms through which TAOK2 regulates ciliary growth and signaling will be determined using super-resolution imaging and biochemical approaches (Aim2). The contribution of TAOK2 in the ciliary defects observed in patient-derived NPCs from 16p11.2 deletion and duplication carriers will be determined through application of quantitative proteomics and genome editing techniques (Aim3). Utilizing a combination of innovative approaches and human disease relevant model systems, we seek to understand how TAOK2 kinase signaling mediates neural stem cell development, and how perturbations in its signaling pathways contribute to the clinical neuropathology of 16p11.2 CNV. !
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0.955 |