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Principal Investigator: Pamela Robey
Organization: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
Fiscal Year: 2021
Award: $865,069
Funding agency: National Institute of Neurological Disorders and Stroke
During the last fiscal year, the NIH SCCF has made progress in a number of areas as highlighted below.
Characterization of RSet nave-like pluripotent stem cells:
One of the essential properties of human pluripotent stem cells (hPSCs) is the ground or nave pluripotent state that is primarily established in mouse embryonic stem cells (mESCs). Thus far, more than five well-established research groups have reported the existence of human nave pluripotency. However, cell growth conditions and standards have not been defined or established. With Dr. Barbara Mallon, a former SCU member, we have characterized several nave-like pluripotent stem cell lines (NLPs) in terms of their growth patterns, signaling requirements, gene expression profiles, and global methylation. Our defined NLP growth conditions (RSet) may have significant benefits for hPSC growth, downstream differentiation, genetic engineering, disease modeling, and drug discovery.
Establishment of a meta-analysis platform for stem cell research:
In collaboration with Dr. Kory Johnson and Dr. Yang Fann of the NINDS Bioinformatics IT Group, we established an integrated multivariate meta-analysis platform, significantly improving the predictive value of the current meta-analysis. Accordingly, we were able to reveal various similarities between some nave-like hPSCs and their human and mouse in vitro counterparts. Moreover, we also showed numerous fundamental inconsistencies between diverse nave-like states, which are likely attributed to inter-laboratory protocol differences. This meta-analysis platform is suitable for a broad range of biomedical applications, particularly for stem and cancer cell research.
Unraveling SARS-CoV-2 infectivity and predicting therapeutic responses: roles of complex stem-cell organoids:
The COVID-19 pandemic is a rapidly evolving and life-threatening situation caused by the SARS-CoV-2 coronavirus. In collaboration with Dr. Kyeyoon Park and Dr. Jason Spence (University of Michigan), we highlighted the emerging roles of complex organoids derived from human pluripotent stem cells and adult stem cells in SARS-CoV-2 cell-entry assays, COVID-19 drug, and in antibody and vaccine development. We provided new insights into the utility of these organoid assays in clinical management of COVID-19 patients during the pandemic, thereby facilitating targeted prevention in vulnerable populations and a precision-and-effective therapy for individual COVID-19 patients.
Generation of disease models:
We have used the CRISPR/cas9 system to generate cell clones with disruptions in Protein deglycase DJ-1, Parkin,TBK1 and TREM2. Protein deglycase DJ-1, also known as Parkinsons disease protein 7, is a protein which in humans is encoded by the PARK7 gene. Defects in this gene are the cause of autosomal recessive early-onset Parkinson disease. Parkin, also known as Parkinson disease protein 2, is a E3 ubiquitin ligase that plays a critical role in ubiquitination. Parkin recognizes proteins on the outer membrane of mitochondria upon cellular insult and mediates the clearance of damaged mitochondria via autophagy and proteasomal mechanisms. Parkin also enhances cell survival by suppressing both mitochondria-dependent and independent apoptosis. Mutations are associated with mitochondrial dysfunction, leading to neuronal death in autosomal recessive juvenile Parkinson disease. TBK1 (TANK-binding kinase 1) is an enzyme with serine/threonine kinase activity, known for its role in innate immunity antiviral response, but it also regulates cell proliferation, apoptosis, autophagy, and anti-tumor immunity. Loss-of-function mutations of TBK1 are known to cause frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), often combined with memory deficits early in the disease course. TREM2 (triggering receptor expressed on myeloid cells-2) is a surface receptor belonging to the immunoglobulin superfamily. TREM2 is involved in several cellular functions such as cell survival, phagocytosis, pro-inflammatory cytokine production, and cytoskeletal rearrangement. Loss of function mutations in the TREM2 gene are associated with Nasu-Hakola disease, a dementia with bone cysts. Rare heterozygous variants of TREMs are associated with increased risk of developing late-onset Alzheimer disease.
Sjogren syndrome is a chronic autoimmune disorder that causes insufficient moisture production in certain glands of the body, including the saliva-producing and tear-producing glands. To study the pathophysiology of Sjogrens syndrome it is imperative to develop an in vitro model system which can reconstitute 3D salivary glands. As an initial step toward building models, induced pluripotent stem cells were generated from salivary epithelial cells of Sjogren syndrome patients in collaboration with Dr. Youngmi Ji at NIDCR.
Glial differentiation:
Glial cells are increasingly recognized to play a critical role in the development and function of the brain. Moreover, glial cell dysfunction has been shown to contribute to various neurological disorders. Therefore, understanding the function of glial cells under normal as well as pathological conditions is important. To develop optimal protocols for glial differentiation and to provide cells for each glial lineage, human pluripotent stem cells (hPSCs) were employed as a source. As the first target, astrocytes were generated from hPSCs. To do this, hPSCs were first differentiated to neural precursor cells (NPCs), and NPCs were differentiated into astrocyte precursors. The astrocyte precursors were then matured to astrocytes.
Microglia are cells of mesodermal/mesenchymal origin that migrate into the CNS to become resident macrophages within the unique brain microenvironment. They play crucial role in the healthy brain as regulators of synaptic functions and phagocytosis. In disease, they play a crucial role in neurodegenerative and neuro-inflammatory conditions. To generate microglia from pluripotent stem cells, hiPSCs were first differentiated into the hematopoietic lineage. The hematopoietic progenitors are then further differentiated into microglia lineage.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><3-D><3-Dimensional><3D><ALS and FTD><ALS and FTLD><Amentia><Amyotrophic lateral sclerosis and frontotemporal degeneration><Amyotrophic lateral sclerosis and frontotemporal dementia><Amyotrophic lateral sclerosis and frontotemporal lobar dementia><Anti-Viral Response><Antibodies><Antiviral Response><Apoptosis><Apoptosis Pathway><Area><Assay><Astrocytes><Astrocytus><Astroglia><Autoimmune Diseases><Autophagocytosis><Bio-Informatics><Bioassay><Bioinformatics><Biologic Assays><Biologic Models><Biological Assay><Biological Models><Blood Precursor Cell><Bone Cysts><Brain><Brain Nervous System><COVID crisis><COVID epidemic><COVID infected patient><COVID pandemic><COVID patient><COVID positive patient><COVID-19><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 infected patient><COVID-19 pandemic><COVID-19 patient><COVID-19 positive patient><COVID-19 public health crisis><COVID-19 virus><COVID19><COVID19 crisis><COVID19 epidemic><COVID19 global health crisis><COVID19 global pandemic><COVID19 health crisis><COVID19 pandemic><COVID19 patient><COVID19 positive patient><COVID19 public health crisis><COVID19 virus><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CV-19><CV19><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Culture Techniques><Cell Differentiation><Cell Differentiation process><Cell Function><Cell Growth in Number><Cell Line><Cell Multiplication><Cell Process><Cell Proliferation><Cell Signaling><Cell Survival><Cell Viability><Cell physiology><CellLine><Cells><Cellular Expansion><Cellular Function><Cellular Growth><Cellular Physiology><Cellular Process><Cellular Proliferation><Chronic><Clinic><Clinical Management><Clone Cells><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><CoV-2><CoV2><Collaborations><Complex><Coronaviridae><Coronavirus><DJ-1><DJ1><Defect><Dementia><Derivation><Derivation procedure><Development><Disease><Disease model><Disorder><Drugs><Dysfunction><E3 Ligase><E3 Ubiquitin Ligase><ES cell><Educational process of instructing><Encephalon><Enzyme Gene><Enzymes><Epithelial Cells><FLJ11330><FTD/ALS><FTLD-ALS><Functional disorder><Gene Expression><Generalized Growth><Generations><Genes><Genetic Alteration><Genetic Change><Genetic Engineering><Genetic Engineering Biotechnology><Genetic Engineering Molecular Biology><Genetic defect><Genome Instability><Genomic Instability><Gland><Glia><Glial Cells><Glial Differentiation><Goals><Growth><Hematopoietic><Hematopoietic Progenitor Cells><Hematopoietic stem cells><Hortega cell><Human><Immune Globulins><Immunoglobulins><In Vitro><Individual><Inflammatory><Innate Immunity><Intracellular Communication and Signaling><Juvenile Parkinson Disease><Juvenile Parkinsonism><Kolliker's reticulum><Laboratories><Late Onset Alzheimer Disease><Life><Malignant Cell><Mediating><Medication><Memory Deficit><Memory impairment><Mentors><Mesenchymal><Mesoderm Cell><Mesodermal Cell><Meta-Analysis><Methods><Mice><Mice Mammals><Michigan><Microglia><Mission><Mitochondria><Model System><Modern Man><Monitor><Murine><Mus><Mutation><Mφ><NF-KB-Activating Kinase><NF-KB-Activating Kinase NAK><NF-Kb-Activating Kinase Gene><NIDCR><NIDR><NIH><NINDS><National Institute of Dental Research><National Institute of Dental and Craniofacial Research><National Institute of Neurological Diseases and Stroke><National Institute of Neurological Disorders and Stroke><National Institutes of Health><Native Immunity><Natural Immunity><Nerve Degeneration><Nervous System Diseases><Neural Stem Cell><Neuroglia><Neuroglial Cells><Neurologic Disorders><Neurological Disorders><Neuron Degeneration><Non-Specific Immunity><Non-neuronal cell><Nonneuronal cell><Nonspecific Immunity><Oncogene DJ1><Organoids><Outer Mitochondrial Membrane><PARK2 protein><PARK7><PARK7 gene><Paralysis Agitans><Parkin><Parkinson><Parkinson Disease><Parkinson disease 2 protein><Parkinson's disease><Parkinsons disease><Pathologic><Patients><Pattern><Phagocytosis><Pharmaceutic Preparations><Pharmaceutical Preparations><Physiopathology><Play><Pluripotent Stem Cells><Pre-clinical Drug Testing/Development><Preclinical Drug Development><Preclinical Drug Testing/Development><Prediction of Response to Therapy><Predictive Value><Prevention><Primary Parkinsonism><Process><Production><Progenitor Cells><Programmed Cell Death><Property><Protein-Serine Kinase><Protein-Serine-Threonine Kinases><Protein-Threonine Kinase><Proteins><Protocol><Protocols documentation><Quality Control><Receptor Protein><Recombinant DNA Technology><Reporting><Research><Risk><Role><SARS corona virus 2><SARS-CoV-2><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 infected patient><SARS-CoV-2 pandemic><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV2><SARS-CoV2 epidemic><SARS-CoV2 pandemic><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Saliva><Salivary><Salivary Glands><Salivary Glands Head and Neck><Sampling><Serine Kinase><Serine-Threonine Kinases><Serine/Threonine Protein Kinase Gene><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome related corona virus 2><Sicca Syndrome><Side><Signal Transduction><Signal Transduction Systems><Signaling><Simple Bone Cyst><Sjogren's Syndrome><Sjogrens><Sjögren Syndrome><Solitary Bone Cyst><Solitary Cysts><Source><Stem Cell Research><Strains Cell Lines><Subcellular Process><Surface><System><T2K><TANK Binding Kinase TBK1><TANK-binding kinase 1><TBK1><TBK1 gene><TREM2><TREM2 gene><Teaching><Threonine Kinase><Tissue Growth><Triggering Receptor Expressed on Myeloid Cells 2><Tumor Immunity><Ubiquitilation><Ubiquitin Protein Ligase><Ubiquitin-Protein Ligase Complexes><Ubiquitin-Protein Ligase E3><Ubiquitination><Ubiquitinoylation><Unicameral Bone Cyst><United States National Institutes of Health><Universities><Variant><Variation><Vulnerable Populations><Wuhan coronavirus><Yang><adult stem cell><amyotrophic lateral sclerosis and frontotemporal lobar degeneration><amyotrophic lateral sclerosis/ftd><anti-tumor immunity><antitumor immunity><astrocytic glia><autoimmune condition><autoimmune disorder><autophagy><biological signal transduction><blood stem cell><cancer cell><cancer immunity><cell community><cell culture><cell growth><cellular community><clinical center><corona virus><corona virus disease 2019><corona virus disease 2019 epidemic><corona virus disease 2019 pandemic><coronavirus disease 2019><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global 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loss><neuronal progenitor><neuronal progenitor cells><neuronal stem cells><neuroprogenitor><ontogeny><pandemic><pandemic disease><parkin gene/protein><pathophysiology><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><perivascular glial cell><pluripotency><pre-clinical drug development><predict therapeutic response><predict therapy response><predict treatment response><progenitor Cell growth><receptor><reconstitute><reconstitution><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 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