Our lab is focused on high content drug screening, assay development and the combination of electrophysiological, imaging and genetic assays to deeply interrogate human induced pluripotent stem cell (hiPSC)-derived neurons. Our team has developed novel techniques for screening and analyzing 3D organoids developed from hiPSCs to model cortical development in autism.

My graduate work in the lab of Dr. Scott Thompson at the University of Maryland, School of Medicine focused on single cell approaches to study the relationship between synaptic morphology and synaptic plasticity at the tripartite synapse in the hippocampus. To do this, I used a combination of live-cell imaging and advanced techniques in electrophysiology, including single-synapse microphotolysis combined with gene gun transfection in organotypic hippocampal slice cultures. We found that the expression of the ephrinA4 receptor on astrocytes affected their morphology and subsequent synaptic activity.

As a postdoctoral fellow at The National Institutes of Health in Dr. Dax Hoffman’s lab, I focused on real-time imaging techniques such as in vivo two-photon microscopy, FRAP, in vivo single-cell electroporation and calcium imaging. This work included the creation and use of track-tracing trans-synaptic rabies virus and Sindbis virus designed to express GFP-tagged Kv4.2 channels. By using two-photon imaging at single dendrites we were able to demonstrate that Kv4.2 channels are differentially trafficked at different regions along the apical dendrite of neurons during basal activity and upon stimulation in a PKA-dependent manner.

Because of my interest in human induced pluripotent stem cells (iPSCs) and their application in disease modeling, I joined Dr. Scott Noggle at The New York Stem Cell Foundation and developed a number of protocols surrounding the use of a three-dimensional iPSC culture system called a serum-free embryoid body (SFEB). Using SFEBs, we were able to perform electrophysiology combined with calcium imaging as well as develop a protocol to create interneurons that shared transcriptional histories and functional properties with those that were born in the caudal ganglionic eminence.

My approach at The Hussman Institute for Autism involves leveraging the sum of these experiences to develop a robust human cellular model of autism suitable for preclinical drug discovery and therapeutic development.

Kilander MBC, Wang C-H, Chang C-H, Nestor JE, Herold K, Tsai J-W, Nestor MW, Lin Y-C (2018) A rare human CEP290 variant disrupts the molecular integrity of the primary cilium and impairs Sonic Hedgehog machinery. Scientific Reports 8.

Lunden JW, Durens M, Phillips AW, Nestor MW (2018) Cortical interneuron function in autism spectrum condition. Pediatric Research 85:146–154.

Derosa BA, Hokayem JE, Artimovich E, Garcia-Serje C, Phillips AW, Booven DV, Nestor JE, Wang L, Cuccaro ML, Vance JM, Pericak-Vance MA, Cukier HN, Nestor MW*, Dykxhoorn DM (2018) Convergent Pathways in Idiopathic Autism Revealed by Time Course Transcriptomic Analysis of Patient-Derived Neurons. Scientific Reports 8. *Co-senior author.

Artimovich, E., Jackson, R. K., Kilander, M. B. C., Lin, Y. C., & Nestor, MW. (2017). PeakCaller: an automated graphical interface for the quantification of intracellular calcium obtained by high-content screening. BMC Neurosci, 18(1), 72. doi:10.1186/s12868-017-0391-y

Flaherty, E., Deranieh, R. M., Artimovich, E., Lee, I. S., Siegel, A. J., Levy, D. L, *Nestor MW, *Brennand, K. J. (2017). Patient-derived hiPSC neurons with heterozygous CNTNAP2 deletions display altered neuronal gene expression and network activity. NPJ Schizophr, 3(1), 35. doi:10.1038/s41537-017-0033-5 *Co-senior authors.

Phillips AW, Nestor JE, Nestor MW. Developing HiPSC Derived Serum Free Embryoid Bodies for the Interrogation of 3-D Stem Cell Cultures Using Physiologically Relevant Assays. J Vis Exp. 2017 Jul 20;(125). doi: 10.3791/55799.

Xu J, Hartley BJ, Kurup P, Phillips A, Topol A, Xu M, Ononenyi C, Foscue E, Ho SM, Baguley TD, Carty N, Barros CS, Müller U, Gupta S, Gochman P, Rapoport J, Ellman JA, Pittenger C, Aronow B, Nairn AC, Nestor MW, Lombroso PJ, Brennand KJ (2016) Inhibition of STEP61 ameliorates deficits in mouse and hiPSC-based schizophrenia models. Mol Psychiatry.

Nestor MW, Phillips AW, Artimovich E, Nestor JE, Hussman, JP, Blatt GJ (2015) Human Induced Pluripotent Stem Cells and Autism Spectrum Disorder: Emerging Technologies. Autism Research Oct 1. doi: 10.1002/aur.1570.

Nestor MW, Artimovich E, Wilson RL (2015) The Ethics of Gene Editing Technologies in Human Stem Cells. Ethics in Biology, Engineering and Medicine: An International Journal DOI: 10.1615/EthicsBiologyEngMed.2015013732

Nestor MW, Jacob S, Sun B, Pre D, Sproul AA, Hong SI, Woodard C, Zimmer M, Chinchalongporn V, Arancio O, Noggle SA (2015) Characterization of a subpopulation of developing cortical interneurons from human iPSCs within serum free embryoid bodies. American Journal of Physiology Cell Physiology: C209-C219. *[Journal “Images of the Week” Selection]

Prè D, Nestor MW, Sproul AA, Jacob S, Koppensteiner P, et al. (2014) A Time Course Analysis of the Electrophysiological Properties of Neurons Differentiated from Human Induced Pluripotent Stem Cells (iPSCs). PLoS ONE 9(7): e103418. doi:10.1371/journal.pone.0103418.

Woodard, CM, Campos BA , Kuo SH, Nirenberg MJ, Nestor MW, Zimmer M, Mosharov EV, Sulzer D, Zhou H, Paull D, Clark L, Schadt EE, Sardi SP, Rubin L, Eggan K, Brock M, Lipnick S, Rao M, Chang S, Li A, Noggle SA (2014). iPSC-Derived Dopamine Neurons Reveal Differencesbetween Monozygotic Twins Discordant for Parkinson’s Disease. Cell Rep 9(4): 1173-1182. *[Cell Reports Cover Selection]

Yamada M, Johannesson B, Sagi I, Cole-Burnett L, Kort DH, Prosser RW, Paull D, Nestor MW, Freeby M, Greenberg E, Goland RS, Leibel RL, Solomon SL, Benvenisty N, Sauer MV, Egli D. (2014) Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells. Nature 10.1038/nature13287. *[Article featured in “The Scientist”]

Sproul AA, Jacob S, Pre D, Kim SH, Nestor MW, et al. (2014) Characterization and Molecular Profiling of PSEN1 Familial Alzheimer’s Disease iPSC-Derived Neural Progenitors. PLoS ONE 9(1): e84547. doi:10.1371/journal.pone.0084547.

Nestor MW, Noggle SA (2013). Standardization of human stem cell pluripotency using scorecards and bioinformatics. Stem Cell Research & Therapy 4:37.

Nestor MW, Paull D, Jacob S, Sproul AA, Alsaffar A, Campos BA, Noggle SA (2013) Differentiation of serum-free embryoid bodies from human induced pluripotent stem cells into networks. Stem Cell Res 10:454-463. *[Article covered by “Science Today” and “The Source” (Canada)]

Marshall C, Hua H, Shang L, Ding BS, Zito G, de Peppo GM , Wang GK, Douvaras P, Sproul AA., Paull D, Fossati V, Nestor MW, McKeon D, Smith KA. Solomon SL (2012). The Sixth Annual Translational Stem Cell Research Conference of the New York Stem Cell Foundation. Annals of the New York Academy of Sciences.

Nestor MW, Hoffman DA (2012). Aberrant dendritic excitability: a common pathophysiology in CNS disorders affecting memory? Mol Neurobiol 45:478-487.

Nestor MW, Hoffman DA (2011). Differential cycling rates of Kv4.2 channels in proximal and distal dendrites of hippocampal CA1 pyramidal neurons. Hippocampus, 21: doi: 10.1002/hipo.20899.

Nestor MW, Cai X, Bloch RJ, Thompson SM (2011). beta-spectrin regulates the morphological and functional dynamics of dendritic spines via its actin-binding domain and interaction with Rac3. PLOS One. PLoS ONE 6(1): e16197. doi:10.1371/journal.pone.0016197.

Nestor MW, Mok L, Tulapurkar ME, & Thompson SM (2007). Plasticity of Neuron-Glial Interactions Mediated by Astrocytic EphARs. Journal of Neuroscience, 27(47), 12817-12828. *[Featured article in “This Week” in The Journal of Neuroscience]

Mattison H, Nestor MW, Thompson SM (2007) LTD in cerebellar Purkinje cells: what do spines have to do with it? Cellscience Reviews: July (4): 26-36.

Thompson SM, Mattison H, Nestor MW (2005). Morphological correlates of long-term potentiation and depression. Cellscience Reviews: Oct. (2): 58-78.

Parenté F, Nestor MW, Stouffer E, Wingrat J, Hiob T. (2003). Measuring invisible cognitions: A technique and applications. Cognitive Technology, 8(2), 37-41.