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Principal Investigator: HERBERT M LACHMAN
Organization: ALBERT EINSTEIN COLLEGE OF MEDICINE
Fiscal Year: 2024
Award: $252,000
Funding agency: National Institute of Mental Health
Program Director/Principal Investigator (Lachman, Herbert M)
Abstract
Mutations in PPM1D (Protein Phosphatase Magnesium-Dependent 1 Delta), which codes for a member of the
protein serine-threonine phosphatase family, have recently been found in individuals with Jansen-De Vries
Syndrome (JdVS), a condition characterized by intellectual disability (ID), severe anxiety, oppositional
behavior, attention problems, high pain threshold, restricted eating, and gastrointestinal problems. So far,
several dozen children with JdVS have been reported in the literature. Remarkably, each has a de novo
germline nonsense or frameshift mutation in exon 5 or 6 that predicts the formation of a protein truncated at the
C-terminal end. The phosphatase domain is spared. A key question is whether the truncating mutations result
in gain- or loss- of PPM1D function in neurons and other brain cells. PPM1D is also a tumor suppressor gene
that is over-expressed in a variety of cancer subtypes, contributing to malignant transformation through its
action as a negative regulator of the p53-mediated DNA repair pathway. In cancers, somatic, truncating
mutations in exons 5 and 6 like those found in JdVS, but somatic in origin, result in gain-of-function (GOF)
effects because PPM1D degradation is reduced. Yet, loss-of-function (LOF) variants in the catalytic domain
have also been found in some cancers, suggesting that the effects of PPM1D on cellular function could be
context-dependent. Whether the germline truncating mutations in children with JdVS cause GOF or LOF is a
critical question to address because PPM1D inhibitors being developed by cancer researchers could
potentially have therapeutic value in JdVS if the former is correct. On the other hand, if they cause LOF, drugs
targeting over-phosphorylated downstream targets caused by reduced PPM1D phosphatase activity, would be
a more feasible approach. To address these fundamental questions, we have established an induced
pluripotent stem cell (iPSC) model for JdVS using patient-specific and CRISPR-engineered lines, each
containing exon 5 or exon 6 truncating mutations. Preliminary RNA-seq and proteomics experiments on
glutamatergic neurons derived from iPSCs identified a few functional signatures, including deficits in neurite
outgrowth, altered chromatin architecture, and calmodulin kinase 2 (CAMK2) phosphorylation that suggest a
GOF effect. However, molecular studies need to be repeated and expanded in additional lines to firmly
establish these as legitimate phenotypes, and to identify novel makers that can be used to both test the GOF
hypothesis and identify downstream targets for therapeutic intervention. This will be accomplished by a multi-
OMICs approach on glutamatergic neurons that includes proteomics, phosphoproteomics, RNA-seq, and
ATAC-seq. Considering the uniqueness of the PPM1D mutations in the development of JdVS, we also
hypothesize that truncated PPM1D proteins have powerful effects on cellular function that could make them
resistant to typical PPM1D inhibitors. To test these hypotheses, we will knockout either the mutant allele or wild
type PPM1D allele by introducing a null mutation using CRISPR-Cas9 editing, after which, rescue of
dysregulated cellular and molecular phenotypes will be assessed. The ability of small molecule PPM1D
inhibitors to rescue these phenotypes will also be evaluated. The experiments will show whether JdVS
associated PPM1D variants have a GOF effect on glutamatergic neurons, and whether generalized reduction
in PPM1D phosphatase activity or targeted inhibition of the truncated protein will be suitable targets for
therapeutic intervention.
Terms: <0-11 years old><1st trimester><3-D structure><3-dimensional structure><3D structure><ASD><ATAC sequencing><ATAC-seq><ATACseq><Achievement><Achievement Attainment><Address><Alleles><Allelomorphs><Anti-Oncogenes><Antioncogene Protein p53><Antioncogenes><Appearance><Architecture><Assay for Transposase-Accessible Chromatin using sequencing><Attention><Autism><Autistic Disorder><Autoimmune><B.1.617.2><Behavior><Biologic Models><Biological><Biological Models><Blood><Blood Reticuloendothelial System><Blood leukocyte><Brain><Brain Nervous System><C-terminal><CRISPR><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Ca(2+)-Calmodulin Dependent Protein Kinase><CaMK><Calcium/calmodulin-dependent protein kinase><Calmodulin-Dependent Protein Kinases><Calmodulin-Kinase><Cancer Suppressor Genes><Cancers><Cas nuclease technology><Catalytic Core><Catalytic Domain><Catalytic Region><Catalytic Site><Catalytic Subunit><Cell Body><Cell Function><Cell Line><Cell Physiology><Cell Process><CellLine><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cellular Tumor Antigen P53><Cerebrum><Child><Child Youth><Childhood><Children (0-21)><Chromatin><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats approach><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><Code><Coding System><Communities><DNA Repair Pathway><Delta variant><Development><Diameter><Disease><Disorder><Dll4 protein><Doctor of Medicine><Drug Targeting><Drugs><Early Infantile Autism><Early Placental Phase><Eating><Emerogenes><Encephalon><Engineering><Engineering / Architecture><Exons><Family><First Pregnancy Trimester><First Trimester><Food Intake><Frame Shift Mutation><Frameshift Mutation><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Germ Lines><Glutamates><Impairment><Individual><Induced pluripotent stem cell derived neurons><Infantile Autism><Inflammatory><Intellectual disability><Intellectual functioning disability><Intellectual limitation><Investigators><Kanner's Syndrome><Knock-out><Knockout><Knowledge><L-Glutamate><Leukocytes><Leukocytes Reticuloendothelial System><Literature><M.D.><Magnesium><Malignant><Malignant - descriptor><Malignant Neoplasms><Malignant Tumor><Marrow leukocyte><Mediating><Medication><Mg element><Microscopic><Model System><Modernization><Molecular><Molecular Analysis><Mutate><Mutation><Nerve Cells><Nerve Unit><Neural Cell><Neurites><Neurocyte><Neuron from iPSC><Neuron from induced pluripotent stem cells><Neurons><Non-Polyadenylated RNA><Nonsense Mutation><Obsessive-Compulsive Disorder><Obsessive-Compulsive Neurosis><Onco-Suppressor Genes><Oncogenes-Tumor Suppressors><Oncoprotein p53><Organoids><Outcome><P53><PANDAS><Pain Threshold><Pain Tolerance Level><Pathological anxiety><Patients><Pediatric Acute-Onset Neuropsychiatric Syndrome><Pediatric Autoimmune Neuropsychiatric Disorder><Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcus Infections><Pharmaceutical Preparations><Phenotype><Phosphatases><Phosphohydrolases><Phosphomonoesterases><Phosphoprotein P53><Phosphoprotein Phosphatase><Phosphoprotein Phosphatase-2C><Phosphoprotein Phosphohydrolase><Phosphoprotein pp53><Phosphoproteins><Phosphoric Monoester Hydrolases><Phosphorylation><Principal Investigator><Protein Phosphatase C><Protein Phosphatase Gene><Protein Phosphatase-1><Protein Phosphatase-2A><Protein Phosphorylation><Protein Serine/Threonine Phosphatase><Protein Serine/Threonine Phosphatase Gene><Protein TP53><Protein Truncation><Protein phosphatase><Proteins><Proteomics><QOL><Quality of life><RNA><RNA Gene Products><RNA Seq><RNA sequencing><RNAseq><Reading Frame Shift Mutation><Recessive Oncogenes><Reporting><Research><Research Personnel><Research Resources><Researchers><Resistance><Resources><Ribonucleic Acid><SARS-CoV-2 B.1.617.2><SARS-CoV-2 delta><Serine/Threonine Specific Protein Phosphatase Gene><Serine/Threonine-Specific Protein Phosphatase Gene><Stem Cell Development><Strains Cell Lines><Subcellular Process><Symptoms><Syndrome><TP53><TP53 gene><TRP53><Testing><Therapeutic><Therapeutic Effect><Therapeutic Intervention><Transcript><Tumor Protein p53><Tumor Protein p53 Gene><Tumor Suppressing Genes><Tumor Suppressor Genes><Variant><Variation><White Blood Cells><White Cell><anti-cancer research><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><autism spectral disorder><autism spectrum disorder><autistic individuals><autistic people><autistic spectrum disorder><biologic><brain cell><calcium-calmodulin-dependent PK><calcium-calmodulin-dependent PK type II><calmodulin dependent protein kinase><cancer research><cancer sub-types><cancer subtypes><cell type><cerebral><cultured cell line><delta protein><developmental><differential expression><differentially expressed><drug/agent><experiment><experimental research><experimental study><experiments><feeding><fetal><gain of function><gastrointestinal><gene editing method><gene editing methodology><gene editing strategy><gene editing techniques><gene null><gene-editing approach><genome mutation><glutamatergic><iPS><iPS neurons><iPSC><iPSC derived-neurons><iPSC technology><iPSCs><improved><individuals on the autism spectrum><individuals on the spectrum><individuals with ASD><individuals with autism><individuals with autism spectrum disorder><induced pluripotent cell><induced pluripotent stem cell><induced pluripotent stem cell neurons><induced pluripotent stem cell technology><induced pluripotent stem cells derived from patients><induced pluripotent stem cells from patients><inducible pluripotent stem cell><inhibitor><intellectual and developmental disability><interest><intervention therapy><kids><limited intellectual functioning><loss of function><loss of function mutation><malignancy><member><microtubule associated protein 2 kinase><millimeter><molecular biomarker><molecular marker><molecular phenotype><multiomics><multiple omics><mutant><neoplasm/cancer><neuronal><neurons derived from induced pluripotent stem cells><neuropsychiatric><neuropsychiatry><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><non-sense mutation><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><null mutation><oncosuppressor gene><overexpress><overexpression><p53 Antigen><p53 Genes><p53 Tumor Suppressor><pain tolerance><panomics><patient derived human iPS><patient derived human iPSC><patient derived human induced pluripotent stem cell><patient derived iPS><patient derived iPSC><patient derived induced pluripotent cells><patient derived induced pluripotent stem cells><patient-derived pluripotent stem cells><pediatric><pediatric acute neuropsychiatric syndrome><people on the autism spectrum><people with ASD><people with autism><people with autism spectrum disorder><pharmacologic><phenotypic biomarker><phenotypic marker><phospho-proteomics><phosphoproteomics><progenitor cell development><progenitor cell model><progenitor development><progenitor model><programs><protein kinase II><protein p53><public health relevance><resistant><serine-threonine phosphatase><severe acute respiratory syndrome coronavirus 2 B.1.617.2><severe anxiety><small molecule><stem and progenitor cell development><stem and progenitor cell model><stem cell based model><stem cell derived model><stem cell model><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><three dimensional structure><tool><transcriptional differences><transcriptome sequencing><transcriptomic sequencing><transcriptomics><white blood cell><white blood corpuscle><youngster><δ protein>