Bio

Bio


Postdoctoral researcher with a specialty in molecular biology and genetics and a commitment to helping advance medical science and biotechnology

Background: PhD in Biomedical Science with Advanced Concentration in Genetics and combined 8+ years of laboratory research, providing me with the knowledge and experience necessary to help tackle biomedical challenges

Expertise: gene editing; molecular mechanisms of human disease; cell and mouse models of genetic disease; RNA processing regulation; gene structure, expression, and sequencing

Key Skills: proficiency in a range of cellular and molecular biology techniques, including RNA/DNA isolation, RT-PCR, qPCR, SDS-PAGE, western blotting, immunopurification, immunofluorescence, bacterial culture and cloning, mammalian cell culture, siRNA/shRNA-mediated knockdown, transfection, and protein expression. Experienced in global gene expression analysis including next-generation sequencing, microarray analysis, and high-throughput data analysis.

Goals: contributing to a better understanding of disease pathogenesis, developing targeted therapeutics, and improving biotechnology and genomic medicine.

Professional Education


  • Bachelor of Science, Florida International University (2008)
  • Doctor of Philosophy, University of Florida (2014)

Stanford Advisors


Research & Scholarship

Current Research and Scholarly Interests


Developing novel gene editing therapies for genetic autoimmune diseases, including IPEX and IPEX-like syndromes. Generating humanized mouse models of IPEX syndrome to investigate disease pathogenesis and enable therapeutic development.

Publications

All Publications


  • MBNL Sequestration by Toxic RNAs and RNA Misprocessing in the Myotonic Dystrophy Brain CELL REPORTS Goodwin, M., Mohan, A., Batra, R., Lee, K., Charizanis, K., Gomez, F. J., Eddarkaoui, S., Sergeant, N., Buee, L., Kimura, T., Clark, H. B., Dalton, J., Takamura, K., Weyn-Vanhentenryck, S. M., Zhang, C., Reid, T., Ranum, L. P., Day, J. W., Swanson, M. S. 2015; 12 (7): 1159-1168

    Abstract

    For some neurological disorders, disease is primarily RNA mediated due to expression of non-coding microsatellite expansion RNAs (RNA(exp)). Toxicity is thought to result from enhanced binding of proteins to these expansions and depletion from their normal cellular targets. However, experimental evidence for this sequestration model is lacking. Here, we use HITS-CLIP and pre-mRNA processing analysis of human control versus myotonic dystrophy (DM) brains to provide compelling evidence for this RNA toxicity model. MBNL2 binds directly to DM repeat expansions in the brain, resulting in depletion from its normal RNA targets with downstream effects on alternative splicing and polyadenylation. Similar RNA processing defects were detected in Mbnl compound-knockout mice, highlighted by dysregulation of Mapt splicing and fetal tau isoform expression in adults. These results demonstrate that MBNL proteins are directly sequestered by RNA(exp) in the DM brain and introduce a powerful experimental tool to evaluate RNA-mediated toxicity in other expansion diseases.

    View details for DOI 10.1016/j.celrep.2015.07.029

    View details for Web of Science ID 000360130900010

    View details for PubMedCentralID PMC4545389

  • Loss of MBNL Leads to Disruption of Developmentally Regulated Alternative Polyadenylation in RNA-Mediated Disease MOLECULAR CELL Batra, R., Charizanis, K., Manchanda, M., Mohan, A., Li, M., Finn, D. J., Goodwin, M., Zhang, C., Sobczak, K., Thornton, C. A., Swanson, M. S. 2014; 56 (2): 311-322

    Abstract

    Inhibition of muscleblind-like (MBNL) activity due to sequestration by microsatellite expansion RNAs is a major pathogenic event in the RNA-mediated disease myotonic dystrophy (DM). Although MBNL1 and MBNL2 bind to nascent transcripts to regulate alternative splicing during muscle and brain development, another major binding site for the MBNL protein family is the 3' untranslated region of target RNAs. Here, we report that depletion of Mbnl proteins in mouse embryo fibroblasts leads to misregulation of thousands of alternative polyadenylation events. HITS-CLIP and minigene reporter analyses indicate that these polyadenylation switches are a direct consequence of MBNL binding to target RNAs. Misregulated alternative polyadenylation also occurs in skeletal muscle in a mouse polyCUG model and human DM, resulting in the persistence of neonatal polyadenylation patterns. These findings reveal an additional developmental function for MBNL proteins and demonstrate that DM is characterized by misregulation of pre-mRNA processing at multiple levels.

    View details for DOI 10.1016/j.molcel.2014.08.027

    View details for Web of Science ID 000344484600012

    View details for PubMedID 25263597

  • RNA-protein interactions in unstable microsatellite diseases BRAIN RESEARCH Mohan, A., Goodwin, M., Swanson, M. S. 2014; 1584: 3-14

    Abstract

    A novel RNA-mediated disease mechanism has emerged from studies on dominantly inherited neurological disorders caused by unstable microsatellite expansions in non-coding regions of the genome. These non-coding tandem repeat expansions trigger the production of unusual RNAs that gain a toxic function, which involves the formation of RNA repeat structures that interact with, and alter the activities of, various factors required for normal RNA processing as well as additional cellular functions. In this review, we explore the deleterious effects of toxic RNA expression and discuss the various model systems currently available for studying RNA gain-of-function in neurologic diseases. Common themes, including bidirectional transcription and repeat-associated non-ATG (RAN) translation, have recently emerged from expansion disease studies. These and other discoveries have highlighted the need for further investigations designed to provide the additional mechanistic insights essential for future therapeutic development.

    View details for DOI 10.1016/j.brainres.2014.03.039

    View details for Web of Science ID 000343838400002

    View details for PubMedID 24709120

  • RNA-Binding Protein Misregulation in Microsatellite Expansion Disorders SYSTEMS BIOLOGY OF RNA BINDING PROTEINS Goodwin, M., Swanson, M. S. 2014; 825: 353-388

    Abstract

    RNA-binding proteins (RBPs) play pivotal roles in multiple cellular pathways from transcription to RNA turnover by interacting with RNA sequence and/or structural elements to form distinct RNA-protein complexes. Since these complexes are required for the normal regulation of gene expression, mutations that alter RBP functions may result in a cascade of deleterious events that lead to severe disease. Here, we focus on a group of hereditary disorders, the microsatellite expansion diseases, which alter RBP activities and result in abnormal neurological and neuromuscular phenotypes. While many of these diseases are classified as adult-onset disorders, mounting evidence indicates that disruption of normal RNA-protein interaction networks during embryogenesis modifies developmental pathways, which ultimately leads to disease manifestations later in life. Efforts to understand the molecular basis of these disorders has already uncovered novel pathogenic mechanisms, including RNA toxicity and repeat-associated non-ATG (RAN) translation, and current studies suggest that additional surprising insights into cellular regulatory pathways will emerge in the future.

    View details for DOI 10.1007/978-1-4939-1221-6_10

    View details for Web of Science ID 000350418400011

    View details for PubMedID 25201111

  • Generation of Neural Cells from DM1 Induced Pluripotent Stem Cells As Cellular Model for the Study of Central Nervous System Neuropathogenesis CELLULAR REPROGRAMMING Xia, G., Santostefano, K. E., Goodwin, M., Liu, J., Subramony, S. H., Swanson, M. S., Terada, N., Ashizawa, T. 2013; 15 (2): 166-177

    Abstract

    Dystrophia myotonica type 1 (DM1) is an autosomal dominant multisystem disorder. The pathogenesis of central nervous system (CNS) involvement is poorly understood. Disease-specific induced pluripotent stem cell (iPSC) lines would provide an alternative model. In this study, we generated two DM1 lines and a normal iPSC line from dermal fibroblasts by retroviral transduction of Yamanaka's four factors (hOct4, hSox2, hKlf4, and hc-Myc). Both DM1 and control iPSC clones showed typical human embryonic stem cell (hESC) growth patterns with a high nuclear-to-cytoplasm ratio. The iPSC colonies maintained the same growth pattern through subsequent passages. All iPSC lines expressed stem cell markers and differentiated into cells derived from three embryonic germ layers. All iPSC lines underwent normal neural differentiation. Intranuclear RNA foci, a hallmark of DM1, were detected in DM1 iPSCs, neural stem cells (NSCs), and terminally differentiated neurons and astrocytes. In conclusion, we have successfully established disease-specific human DM1 iPSC lines, NSCs, and neuronal lineages with pathognomonic intranuclear RNA foci, which offer an unlimited cell resource for CNS mechanistic studies and a translational platform for therapeutic development.

    View details for DOI 10.1089/cell.2012.0086

    View details for Web of Science ID 000317041100009

    View details for PubMedID 23550732

  • Muscleblind-like 2-Mediated Alternative Splicing in the Developing Brain and Dysregulation in Myotonic Dystrophy NEURON Charizanis, K., Lee, K., Batra, R., Goodwin, M., Zhang, C., Yuan, Y., Shiue, L., Cline, M., Scotti, M. M., Xia, G., Kumar, A., Ashizawa, T., Clark, H. B., Kimura, T., Takahashi, M. P., Fujimura, H., Jinnai, K., Yoshikawa, H., Gomes-Pereira, M., Gourdon, G., Sakai, N., Nishino, S., Foster, T. C., Ares, M., Darnell, R. B., Swanson, M. S. 2012; 75 (3): 437-450

    Abstract

    The RNA-mediated disease model for myotonic dystrophy (DM) proposes that microsatellite C(C)TG expansions express toxic RNAs that disrupt splicing regulation by altering MBNL1 and CELF1 activities. While this model explains DM manifestations in muscle, less is known about the effects of C(C)UG expression on the brain. Here, we report that Mbnl2 knockout mice develop several DM-associated central nervous system (CNS) features including abnormal REM sleep propensity and deficits in spatial memory. Mbnl2 is prominently expressed in the hippocampus and Mbnl2 knockouts show a decrease in NMDA receptor (NMDAR) synaptic transmission and impaired hippocampal synaptic plasticity. While Mbnl2 loss did not significantly alter target transcript levels in the hippocampus, misregulated splicing of hundreds of exons was detected using splicing microarrays, RNA-seq, and HITS-CLIP. Importantly, the majority of the Mbnl2-regulated exons examined were similarly misregulated in DM. We propose that major pathological features of the DM brain result from disruption of the MBNL2-mediated developmental splicing program.

    View details for DOI 10.1016/j.neuron.2012.05.029

    View details for Web of Science ID 000307417700012

    View details for PubMedID 22884328

    View details for PubMedCentralID PMC3418517

  • Functional characterization of an allatotropin receptor expressed in the corpora allata of mosquitoes PEPTIDES Nouzova, M., Brockhoff, A., Mayoral, J. G., Goodwin, M., Meyerhof, W., Noriega, F. G. 2012; 34 (1): 201-208

    Abstract

    Allatotropin is an insect neuropeptide with pleiotropic actions on a variety of different tissues. In the present work we describe the identification, cloning and functional and molecular characterization of an Aedes aegypti allatotropin receptor (AeATr) and provide a detailed quantitative study of the expression of the AeATr gene in the adult mosquito. Analysis of the tissue distribution of AeATr mRNA in adult female revealed high transcript levels in the nervous system (brain, abdominal, thoracic and ventral ganglia), corpora allata-corpora cardiaca complex and ovary. The receptor is also expressed in heart, hindgut and male testis and accessory glands. Separation of the corpora allata (CA) and corpora cardiaca followed by analysis of gene expression in the isolated glands revealed expression of the AeATr primarily in the CA. In the female CA, the AeATr mRNA levels were low in the early pupae, started increasing 6h before adult eclosion and reached a maximum 24h after female emergence. Blood feeding resulted in a decrease in transcript levels. The pattern of changes of AeATr mRNA resembles the changes in JH biosynthesis. Fluorometric Imaging Plate Reader recordings of calcium transients in HEK293 cells expressing the AeATr showed a selective response to A. aegypti allatotropin stimulation in the low nanomolar concentration range. Our studies suggest that the AeATr play a role in the regulation of JH synthesis in mosquitoes.

    View details for DOI 10.1016/j.peptides.2011.07.025

    View details for Web of Science ID 000302851500027

    View details for PubMedID 21839791

  • Allatostatin-C receptors in mosquitoes PEPTIDES Mayoral, J. G., Nouzova, M., Brockhoff, A., Goodwin, M., Hernandez-Martinez, S., Richter, D., Meyerhof, W., Noriega, F. G. 2010; 31 (3): 442-450

    Abstract

    In the present work we describe the functional and molecular characterization of two Aedes aegypti allatostatin-C receptor paralogs (AeAS-CrA and AeAS-CrB) and provide a detailed quantitative study of the expression of the AS-C receptor genes in an adult insect. The tissue distribution of the two AS-C receptors differed significantly; the mRNA levels of AeAS-CrB in the Malpighian tubules were the highest detected, while transcripts for AeAS-CrA were relatively low in this tissue. In addition, the transcript levels of both receptors were different in the thoracic and abdominal ganglia, corpora allata (CA) and the testis of the male. In the CA, the AeAS-CrB mRNA levels were constant from 0 to 72 h after female emergence, while the AeAS-CrA levels increased at 72 h. To complement the receptor expression studies, we analyzed the tissue specificity for allatostatin-C mRNA in female mosquitoes. Expression was high in abdominal ganglia and brain. Transcript levels of allatostatin-C in the head of females were elevated at eclosion and there were no major changes during the first week of adult life or after blood feeding. Fluorometric Imaging Plate Reader (FLIPR) recordings of calcium transients in HEK293T cells transiently expressing both putative receptors showed that they both responded selectively to allatostatin-C stimulation in the nanomolar concentration range. However, the peptide showed slightly greater affinity for AeAS-CrB than AeAS-CrA. Our studies suggest that some of the pleiotropic effects of allatostatin-C in mosquitoes could be mediated by the different receptor paralogs. Transcriptional regulation of the AS-C receptors may not have a critical role in the changes of CA responsiveness to the peptide that we previously described.

    View details for DOI 10.1016/j.peptides.2009.04.013

    View details for Web of Science ID 000275994100011

    View details for PubMedID 19409436