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Ampa Receptor Dysregulation and Therapeutic Interventions in a Mouse Model of Cdkl5 Deficiency Disorder

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Ampa Receptor Dysregulation and Therapeutic Interventions in a Mouse Model of Cdkl5 Deficiency Disorder University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2018 Ampa Receptor Dysregulation And Therapeutic Interventions In A Mouse Model Of Cdkl5 Deficiency Disorder Madhumita Yennawar University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Neuroscience and Neurobiology Commons Recommended Citation Yennawar, Madhumita, "Ampa Receptor Dysregulation And Therapeutic Interventions In A Mouse Model Of Cdkl5 Deficiency Disorder" (2018). Publicly Accessible Penn Dissertations. 3209. https://repository.upenn.edu/edissertations/3209 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/3209 For more information, please contact [email protected]. Ampa Receptor Dysregulation And Therapeutic Interventions In A Mouse Model Of Cdkl5 Deficiency Disorder Abstract CDKL5 Deficiency Disorder (CDD) is a rare disease that presents as a set of neurological deficits including early-life epilepsy, intellectual disability, and autistic-like behaviors. It results from pathogenic mutations in the gene for cyclin-dependent kinase-like 5 (CDKL5), a protein that is highly expressed in brain. There is no cure for CDD and seizures in this disorder are typically resistant to traditional anti-epileptic drugs, although some patients respond well to cannabidiol. However, underlying mechanisms of what causes hyperexcitability and neurological deficits in CDD is poorly understood. We investigated the novel Cdkl5R59X mouse (R59X), and observed that mutant mice have social interaction and memory deficits, and decreased latency to seizure after administration of pentylenetetrazol. Given the observed behavioral alterations and hyperexcitability in R59X mice, we hypothesized that mutant mice would exhibit underlying molecular and functional alterations in proteins involved in regulating the E:I balance. Indeed, we observed a specific decrease in membrane-bound AMPA receptor (AMPAR) subunit GluA2 as well as decreased GluA2:GluA1 in the hippocampus of R59X mice, suggesting an increase in hippocampal GluA2-lacking AMPARs, which are calcium permeable and have significant olesr in regulating neuronal plasticity and excitability. Indeed, decreased hippocampal GluA2 was accompanied by inward rectification of AMPAR currents in whole-cell patch recordings from hippocampal CA1 neurons, indicative of an increased population of functional GluA2-lacking AMPARs, and elevated early-phase long-term potentiation at Schaffer collateral-CA1 synapses. Finally, we evaluated the therapeutic potential of GluA2-lacking AMPAR blocker IEM-1460 as well as cannabidiol to observe 1) whether blocking increased hippocampal GluA2-lacking AMPARs would attenuate behavioral alterations and hyperexcitability in R59X mice and 2) whether cannabidiol shows therapeutic efficacy in R59X mice similaro t human CDD patients, thus validating R59X mice as a relevant model to test potential therapeutics in CDD. Indeed, IEM-1460 significantly escuedr deficits in social behavior, short-term memory, and latency to seizure while cannabidiol significantly escuedr deficits in short and long-term memory and latency to seizure. These results verify that blocking increased GluA2-lacking AMPARs may be a successful therapeutic strategy in CDD, and that both IEM-1460 and cannabidiol may attenuate hyperexcitability as well as autistic-like behaviors and memory deficits in CDD. Degree Type Dissertation Degree Name Doctor of Philosophy (PhD) Graduate Group Pharmacology First Advisor Frances E. Jensen Keywords AMPA, autism, CDKL5, therapeutic Subject Categories Neuroscience and Neurobiology This dissertation is available at ScholarlyCommons: https://repository.upenn.edu/edissertations/3209 AMPA RECEPTOR DYSREGULATION AND THERAPEUTIC INTERVENTIONS IN A MOUSE MODEL OF CDKL5 DEFICIENCY DISORDER Madhumita Yennawar A DISSERTATION in Pharmacology Presented to the Faculties of the University of Pennsylvania in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy 2018 Supervisor of Dissertation _________________________ Dr. Frances E. Jensen Chair, Department of Neurology Graduate Group Chairperson _______________________ Dr. Julie Blendy Professor of Pharmacology Dissertation Committee Dr. Marc A. Dichter, Emeritus Professor of Neurology Dr. Eric Marsh, Associate Professor of Neurology Dr. Zhaolan Zhou, Associate Professor of Genetics Dr. Edward Brodkin, Associate Professor of Psychiatry AMPA RECEPTOR DYSREGULATION AND THERAPEUTIC INTERVENTIONS IN A MOUSE MODEL OF CDKL5 DEFICIENCY DISORDER COPYRIGHT 2018 Madhumita Yennawar This work is licensed under the Creative Commons Attribution- NonCommercial-ShareAlike 3.0 License To view a copy of this license, visit https://creativecommons.org/licenses/by-nc-sa/3.0/us/ ACKNOWLEDGMENT I would first like to thank Penny, Dustin and Harper Howard, who raised the initial funds to start this project. Thank you for sharing your stories with me and showing me how important CDKL5 research is for you and families like yours. I would also like to thank my advisor, Frances Jensen, for the opportunity to join her lab and work on this exciting and fulfilling project. Frances has been a great mentor, and not only taught me about epilepsy and neurodevelopment, but how to think critically, ask the right questions, and demand more of myself. I would also like to acknowledge Marc Dichter, my committee chair, and the rest of my thesis committee for their ideas, guidance, and encouragement. To the Jensen lab members, you have been the most supportive, fun, and helpful colleagues I could have asked for. In particular, thank you to Yeri Song for sharing puppy videos, always having a solution to my problems, and being an amazing friend. Thank you Jen Chan and Christie Ojiaku for being the most wonderful, supportive, and loving friends from day one of graduate school. Finally, I would like to thank my amazing parents, Neela and Hemant, my superstar sister Sneha, and my incredible partner Chris for their endless love and encouragement. Thank you for reading over my documents, listening to my practice talks, and telling me that you believe in me when I needed it most. iii ABSTRACT AMPA RECEPTOR DYSREGULATION AND THERAPEUTIC INTERVENTIONS IN A MOUSE MODEL OF CDKL5 DEFICIENCY DISORDER Madhumita Yennawar Frances E. Jensen CDKL5 Deficiency Disorder (CDD) is a rare disease that presents as a set of neurological deficits including early-life epilepsy, intellectual disability, and autistic-like behaviors. It results from pathogenic mutations in the gene for cyclin-dependent kinase- like 5 (CDKL5), a protein that is highly expressed in brain. There is no cure for CDD and seizures in this disorder are typically resistant to traditional anti-epileptic drugs, although some patients respond well to cannabidiol. However, underlying mechanisms of what causes hyperexcitability and neurological deficits in CDD is poorly understood. We investigated the novel Cdkl5R59X mouse (R59X), and observed that mutant mice have social interaction and memory deficits, and decreased latency to seizure after administration of pentylenetetrazol. Given the observed behavioral alterations and hyperexcitability in R59X mice, we hypothesized that mutant mice would exhibit underlying molecular and functional alterations in proteins involved in regulating the E:I balance. Indeed, we observed a specific decrease in membrane-bound AMPA receptor (AMPAR) subunit GluA2 as well as decreased GluA2:GluA1 in the hippocampus of R59X mice, suggesting an increase in hippocampal GluA2-lacking AMPARs, which are calcium permeable and have significant roles in regulating neuronal plasticity and excitability. Indeed, decreased hippocampal GluA2 was accompanied by inward iv rectification of AMPAR currents in whole-cell patch recordings from hippocampal CA1 neurons, indicative of an increased population of functional GluA2-lacking AMPARs, and elevated early-phase long-term potentiation at Schaffer collateral-CA1 synapses. Finally, we evaluated the therapeutic potential of GluA2-lacking AMPAR blocker IEM-1460 as well as cannabidiol to observe 1) whether blocking increased hippocampal GluA2- lacking AMPARs would attenuate behavioral alterations and hyperexcitability in R59X mice and 2) whether cannabidiol shows therapeutic efficacy in R59X mice similar to human CDD patients, thus validating R59X mice as a relevant model to test potential therapeutics in CDD. Indeed, IEM-1460 significantly rescued deficits in social behavior, short-term memory, and latency to seizure while cannabidiol significantly rescued deficits in short and long-term memory and latency to seizure. These results verify that blocking increased GluA2-lacking AMPARs may be a successful therapeutic strategy in CDD, and that both IEM-1460 and cannabidiol may attenuate hyperexcitability as well as autistic-like behaviors and memory deficits in CDD. v TABLE OF CONTENTS ACKNOWLEDGMENT ......................................................................................... III ABSTRACT ......................................................................................................... IV LIST OF TABLES ................................................................................................. X LIST OF ILLUSTRATIONS ................................................................................. XI CHAPTER 1- INTRODUCTION ............................................................................. 1 Overview .........................................................................................................................
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