WO2024020572A1 - Methods and compositions for the treatment of ptsd - Google Patents

Methods and compositions for the treatment of ptsd Download PDF

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Publication number
WO2024020572A1
WO2024020572A1 PCT/US2023/070761 US2023070761W WO2024020572A1 WO 2024020572 A1 WO2024020572 A1 WO 2024020572A1 US 2023070761 W US2023070761 W US 2023070761W WO 2024020572 A1 WO2024020572 A1 WO 2024020572A1
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glucocorticoid
ptsd
individual
expression
gene
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PCT/US2023/070761
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French (fr)
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Rachel Yehuda
Kristen Brennand
Daniel John PAULL
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New York Stem Cell Foundation, Inc.
Icahn School Of Medicine At Mount Sinai
The United States Government As Represented By The Department Of Veteran Affairs
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Publication of WO2024020572A1 publication Critical patent/WO2024020572A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/20—Hypnotics; Sedatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5058—Neurological cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
    • G01N2333/723—Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/30—Psychoses; Psychiatry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis

Definitions

  • PTSD Post-Traumatic Stress Disorder
  • agents that modify the glucocorticoid response e.g., glucocorticoid rector antagonists
  • psychedelic agents e.g., glucocorticoid rector antagonists
  • BACKGROUND Post-Traumatic Stress Disorder (PTSD) affects 7-8% of the general population of the United States and approximately 15% of veterans returning from combat. The symptoms can persist for months or decades. Unfortunately, PTSD is often misdiagnosed and left untreated in affected civilian and military individuals, disrupting the quality of their lives, their families, and children. Even when diagnosed, the severity of PTSD progression remains difficult to treat.
  • PTSD diagnosis in monozygotic and dizygotic twins, and genome-wide association studies (GWAS) estimate single nucleotide polymorphism (SNP)-based heritability from 5-30% and identify loci significantly associated with PTSD.
  • SNP single nucleotide polymorphism
  • PBMCs peripheral blood mononuclear cells
  • the present disclosure includes the disclosure that the expression and activity of particular biomarkers (e.g., one or more genes of the glucocorticoid response (e.g., a gene from Table 1 and/or Table 2) in biological samples can be utilized to diagnose, prognose, and treat post-traumatic stress disorder (PTSD) in individuals, and further to select individuals who would benefit from a therapy in which one or more psychedelic agents and/or agents that modify the glucocorticoid response are administered. Accordingly, the present disclosure encompasses methods that utilize genes of the glucocorticoid response for the diagnosis, prognosis, and/or treatment of PTSD.
  • biomarkers e.g., one or more genes of the glucocorticoid response (e.g., a gene from Table 1 and/or Table 2) in biological samples
  • PTSD post-traumatic stress disorder
  • the present disclosure encompasses methods that utilize genes of the glucocorticoid response for the diagnosis, prognosis, and/or treatment of
  • psychedelic agents for use in the treatment of post- traumatic stress disorder (PTSD), wherein the individual has modified expression and/or activity of MYC, PAX6, YY1, LEF1, or NFKB1 relative to a suitable control.
  • the individual has modified expression and/or activity of MYC, PAX6, YY1, and LEF1 relative to a suitable control.
  • the individual has increased expression and/or activity of MYC, increased expression and/or activity of PAX6, increased expression and/or activity of YY1, and/or increased expression and/or activity of LEF1.
  • kits for treating an individual having modified expression and/or activity of MYC, PAX6, YY1, LEF1, or NFKB1 relative to a suitable control for post-traumatic stress disorder comprising administering to the individual a therapeutically effective amount of a psychedelic agent.
  • the individual has modified expression and/or activity of MYC, PAX6, YY1, and LEF1 relative to a suitable control.
  • the individual has increased expression and/or activity of MYC, increased expression and/or activity of PAX6, increased expression and/or activity of YY1, and/or increased expression and/or activity of LEF1.
  • the disclosure provides a method of treating an individual diagnosed with PTSD including administering to the individual diagnosed with PTSD a therapeutically effective amount of a psychedelic agent and/or glucocorticoid receptor antagonist.
  • the individual diagnosed with PTSD is diagnosed with PTSD by a the method including: i) obtaining a biological sample from the individual suspected of being at risk for PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) detecting the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response; and v) diagnosing the individual with PTSD if the expression and/or activity of the one or more gene(s) of the glucocorticoid- induced response are modified relative to a suitable control
  • the disclosure provides a method of preventing PTSD in an individual at risk for PTSD comprising administering to the indivusualg a therapeutically effective amount of a glucocorticoid receptor antagonist and/or a psychedelic agent.
  • an individual at risk for PTSD is identified as an individual at risk for PTSD by the method including: i) obtaining a biological sample from the individual suspected of being at risk for PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) detecting the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response; and v) identifying the individual as at risk for PTSD or if the expression and/or activity of the one or more gene(s) of the glucocorticoid-induced response are modified relative to a suitable control.
  • the disclosure provides a method of treating an individual at risk for PTSD, suffering from one or more symptoms associated with PTSD, and/or diagnosed with PTSD, the method including: i) obtaining a biological sample from the individual suspected of being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) detecting the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response; v) identifying the individual as at risk for PTSD or diagnosing the individual with PTSD when the expression and/or activity of the one or more gene(s) of the glucocorticoid-induced response are modified relative to a suitable control; and vi) administering to the individual identified as at risk for PTSD or diagnosed with PTSD
  • the one or more gene(s) of the glucocorticoid-induced response are increased relative to a suitable control.
  • the one or more gene(s) of the glucocorticoid-induced response are decreased relative to a suitable control.
  • the disclosure provides a method for identifying an individual at risk for PTSD or diagnosed with PTSD, the method including: i) obtaining a biological sample from the individual suspected of being at risk for PTSD or diagnosed with PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) detecting the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response; and v) identifying the individual as at risk for PTSD or diagnosing the individual with PTSD if the expression and/or activity of the one or more gene(s) of the glucocorticoid-induced response are modified relative to a suitable control.
  • the disclosure provides a method of treating an individual at risk for developing PTSD, diagnosed with PTSD, or experiencing one or more symptoms associated with PTSD, the method including administering one more psychedelic agent(s) and/or agent(s) that modify the glucocorticoid-induced response.
  • the one or more agent(s) that modify the glucocorticoid-induced response includes a glucocorticoid receptor antagonist.
  • the one or more agent(s) is administered via a parenteral or a non-parenteral route.
  • the disclosure provides a method for screening compounds that reduce the risk of an individual developing PTSD, reduce the risk of an individual developing one or more symptoms of PTSD, and/or alleviate one or more symptoms of PTSD in an individual, the method including: i) obtaining a biological sample from the individual at risk for PTSD or suffering from PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) contacting the test cell with one or more test agent(s); detecting the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response; and v) if the one or more test agent(s) modifies the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response compared to a suitable control, identifying the test agent as a compound that does reduce
  • the one or more gene(s) of the glucocorticoid-induced response include one or more genes selected from the group consisting of MAN1A2, CD1D, CEP350, DISP1, USP37, NPHP3, GOLGA4, KIAA1109, DKK4, BMI1, NEDD4, NF1, CEACAM19, ZNF235, KRCC1, KCTD16, RP11-664D7.4, C8orf87, ANO1, PACS1, UBQLNL, LRRC56, DPYSL4, HMBS, SNRNP35, TM2D3, C17orf75, GATA5, ZNF443, ZC3H12B, RSF1, KPNA4, C12orf57, ATP6V0B, ANAPC11, KCNK2, OR7D2, NUDT16, ANKRD17, ERGIC3, KLF7, PEAK1, ASAP1, MIAT, SGPP2, RPS11, A1
  • the one or more test agent(s) increases the expression and/or activity of one or more gene(s) of the glucocorticoid- induced response.
  • the one or more gene(s) include one or more genes selected from the group consisting of ZC3H12B, RSF1, ANKRD17, KLF7, PEAK1, ASAP1, MIAT, SGPP2, RPS11, A1L2, CSMD1, PEX6, PDE11A, DCTN3, TIMP3, ZNF587, and combinations thereof.
  • the one or more test agent(s) decreases the expression and/or activity of one or more gene(s) of the glucocorticoid-induced response.
  • the one or more gene(s) include one or more genes selected from the group consisting of KPNA4, C12orf57, ATP6V0B, ANAPC11, KCNK2, OR7D2, NUDT16, ERGIC3, MASP1, IL1RAPL2, MRPL11, RBX1, TMEM98, IFT27, PPIB, TOMM22, CALCB, RNF152, and combinations thereof.
  • the suitable control includes a prior biological sample taken from the same individual, a biological sample from a healthy individual not having a risk for PTSD, a biological sample from an individual not having one or more symptoms associated with PTSD, or a biological sample from an individual that is diagnosed with PTSD that has been treated for PTSD.
  • the biological sample includes blood cells and/or fibroblasts.
  • processing the cell obtained from the biological sample includes dedifferentiating the cell to produce an induced pluripotent stem cell (iPSC).
  • iPSC induced pluripotent stem cell
  • the iPSC is differentiated to produce the test cell.
  • the differentiated iPSC includes an induced neuron or an induced peripheral blood mononuclear cell.
  • the test cell includes a neuron or a peripheral blood mononuclear cell.
  • the neuron is a glutamatergic neuron.
  • the glucocorticoid includes a glucocorticoid receptor agonist.
  • the glucocorticoid receptor agonist is dexamethasone or hydrocortisone.
  • the detecting includes sequencing RNA derived from the biological sample.
  • the detecting includes detecting a transcriptional profile of a glucocorticoid-induced response.
  • the detecting includes assessing epigenetic changes of the one or more gene(s) of the glucocorticoid-induced response.
  • the assessing of epigenetic changes includes performing a chromatin immunoprecipitation assay.
  • the expression and/or activity of the one or more gene(s) of the glucocorticoid-induced response is increased for a first subset of genes and decreased for a second subset of genes to produce a glucocorticoid- induced response signature.
  • the suitable control includes substantially no test agent.
  • the processing includes automated reprogramming of the cell obtained from the biological sample.
  • contacting the test cell with a glucocorticoid is performed for a duration of from about 1 hour to about 96 hours (e.g., about 2 hours to about 95 hours, about 3 hours to about 90 hours, about 4 hours to about 80 hours, about 5 hours to about 70 hours, about 10 hours to about 60 hours, about 20 hours to about 50 hours, or about 30 hours to about 40 hours).
  • the glucocorticoid has a concentration of from about 1 nM to about 10 ⁇ M (e.g., about 10 nM to about 9 ⁇ M, about 50 nM to about 8 ⁇ M, about 100 nM to about 7 ⁇ M, about 1 ⁇ M to about 6 ⁇ M, about 2 ⁇ M to about 5 ⁇ M, about 3 ⁇ M to about 4 ⁇ M).
  • contacting the test cell with one or more test agent(s) is performed for a duration of from about 1 hour to about 96 hours (e.g., about 2 hours to about 95 hours, about 3 hours to about 90 hours, about 4 hours to about 80 hours, about 5 hours to about 70 hours, about 10 hours to about 60 hours, about 20 hours to about 50 hours, or about 30 hours to about 40 hours).
  • the test agent has a concentration of from about 1 nM to about 10 ⁇ M (e.g., about 10 nM to about 9 ⁇ M, about 50 nM to about 8 ⁇ M, about 100 nM to about 7 ⁇ M, about 1 ⁇ M to about 6 ⁇ M, about 2 ⁇ M to about 5 ⁇ M, about 3 ⁇ M to about 4 ⁇ M).
  • the disclosure provides a method of identifying a PTSD- dependent glucocorticoid response gene signature, the method including: i) obtaining a biological sample from an individual suspected of being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD; ii) processing a cell obtained from the biological sample to produce a test cell; iii) contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; iv) detecting the expression and/or activity of a plurality of genes; and v) comparing the expression and/or activity of the plurality of genes with the expression and/or activity of the plurality of genes from a suitable control sample obtained from a healthy individual.
  • the individual is a juvenile.
  • methods of treating an individual at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD comprising: administering to the individual one or more psychedelic agent(s) and/or agent(s) that modify the glucocorticoid-induced response, wherein expression and/or activity of one or more gene(s) of the glucocorticoid-induced response in test cells produced from a biological sample obtained from the individual were modified relative to a suitable control when such test cells were contacted with a glucocorticoid.
  • psychedelic agents for use in a method of treating an individual at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD, wherein expression and/or activity of one or more gene(s) of the glucocorticoid-induced response in test cells produced from a biological sample obtained from the individual were modified relative to a suitable control when such test cells were contacted with a glucocorticoid.
  • agents that modify the glucocorticoid response for use in a method of treating an individual at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD, wherein expression and/or activity of one or more gene(s) of the glucocorticoid-induced response in test cells produced from a biological sample obtained from the individual were modified relative to a suitable control when such test cells were contacted with a glucocorticoid.
  • the present disclosure is based, at least in part, on the surprising finding that glucocorticoid-induced (e.g., to dexamethasone (DEX) and hydrocortisone (HCort)) blood and neuronal responses were significantly enriched for immune response, brain development, and neurodevelopmental disorder genes, with specific upregulation of PTSD-associated genes in neurons only.
  • DEX dexamethasone
  • HCort hydrocortisone
  • the present disclosure is also based, at least in part, on the finding that glucocorticoid hypersensitivity occurred in samples from PTSD cases, with diagnosis- specific effects greatest at low doses, and significantly more robust in neurons than peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • a baseline PTSD diagnosis-specific signature was undetectable in either human neurons or PBMCs.
  • This glucocorticoid-response signature was enriched for transcriptomic patterns observed in post-mortem brain tissue from PTSD cases.
  • FIGs.1A-1E are an experimental schematic and a set of graphs, respectively, showing the transcriptional response to dexamethasone (DEX) in peripheral blood mononuclear cells (PBMCs).
  • FIG.1A is a schematic showing the experimental design. PBMCs from 20 post-traumatic stress disorder (PTSD) cases and 20 combat-exposed controls were treated with DEX for 72 hours and RNAseq was performed.
  • PTSD post-traumatic stress disorder
  • RNAseq was performed.
  • FIG.1C is a set of graphs showing the meta-analysis of expression LogFC (differences observed between vehicle and DEX exposure), which was plotted against -log(P value) for each gene. Gray points indicate significantly differentially expressed genes in the meta-analysis.
  • FIG.1D is a set of graphs showing Module eigengene (ME) values from modules identified by weighted gene co-expression network analysis (WGCNA) were correlated with increasing DEX concentrations. Top correlated modules with DEX concentration are shown here (p-values are labeled above each boxplot). Each module was subjected to gene ontology enrichment analysis and the topmost significant enrichment terms and their associated Benjamini-Hochberg adjusted P-values are displayed.
  • FIG.1E is a graph showing the gene set enrichment of DEX-dependent differentially expressed genes across psychiatric disorder and neurodevelopmental gene sets.
  • FIGs.2A-2F are an experimental schematic and a set of graphs, respectively, showing gene expression changes to hydrocortisone (HCort) in human induced pluripotent stem cell (hiPSC)-derived neurons.
  • FIG.2A is a schematic showing the experimental design.
  • hiPSC-derived neurogenin 2 (NGN2) neurons were treated with HCort for 24 hours and RNAseq was performed.
  • FIG.2B depicts a set of photomicrographs of NGN2 neurons stained for neuronal markers NESTIN and MAP2, nucleic marker HOECHST, and green fluorescent protein (GFP) to confirm neuronal identity and morphology across all conditions.
  • GFP green fluorescent protein
  • FIG.2C is a set of graphs showing meta-analyzed differentially expressed genes (DEGs) in response to increasing concentrations of HCort shows robust changes in NGN2-neurons.
  • DEGs differentially expressed genes
  • a comparative analysis of transcriptome-wide log2 fold-changes in response to different concentrations of HCort in NGN2-neurons shows similar responses, indicating a conserved response across all donors to HCort in NGN2-neurons.
  • FIG.2D is a set of graphs showing the meta-analysis of expression LogFC (differences observed between vehicle and HCort exposure), which was plotted against -log(P value) for each gene. Gray points indicate significantly differentially expressed genes in the meta-analysis.
  • FIG.2E is a graph and respective photomicrograph showing the morphological analysis of neurite outgrowth on day 7 in NGN2-neurons showing a dose-dependent decrease in neurite outgrowth with HCort exposure. Representative images of neurite morphology to HCort exposure are shown below.
  • FIG.2F is a graph showing the gene set enrichment of HCort-dependent differentially expressed genes across psychiatric disorder and neurodevelopmental gene sets.
  • FIGs.3A-3B are a set of graphs showing the HCort stimulated co-expression modules in NGN2-neurons.
  • FIG.3A is a set of graphs showing the weighted gene co- expression network analysis (WGCNA), which identified three groups of co-regulated gene modules.
  • WGCNA weighted gene co- expression network analysis
  • FIG.3B is a set of graphs showing the network visualization of protein-protein interactions within modules indicating clusters and network hubs.
  • FIGs.4A-4F are a set of graphs showing the PTSD-positive specific responses to HCort in NGN2-neurons.
  • FIG.4A is a graph showing genes that differ in their response to HCort in PTSD-positive donors compared to PTSD-negative donors, here termed “differential response genes (DRGs),” which were detected in both the 100 nM and 1000 nM dose, indicating PTSD diagnosis-specific responses to HCort.
  • FIG.4B is a set of heat maps showing that significant NGN2-DRGs correctly classify PTSD-positive from PTSD-negative participants using an unsupervised approach.
  • FIG.4C is a set of graphs showing the meta- analysis of Expression LogFC DRGs (differences observed between PTSD-positive and PTSD-negative) was plotted against -log(P value).
  • FIG.4D is a graph showing the gene set enrichment of significant DRGs across psychiatric disorder gene sets (epilepsy, developmental delay, autism spectrum disorder, intellectual disability, schizophrenia, and fragile X messenger ribonucleoprotein (FMRP) targets).
  • FIG.4E is a set of graphs showing the interactive effect of PTSD diagnosis and HCort exposure on gene expression, which are modeled, and three major observed patterns of direction of effect in significantly interactive genes are represented.
  • FIG.4F is a graph showing the logFC of all significantly interactive diagnosis by HCort genes plotted against the P-value of their interaction term, with most significant genes representing those with most significant interactive effects.
  • FIGs.5A-5D are a set of graphs and a schematic, respectively, showing transcription factors driving PTSD hyper-responsivity.
  • FIG.5A is a set of graphs showing PTSD hyper-responsive genes were shown to be enriched for several transcription factor targets.
  • FIG.5B is a schematic of a network visualization of protein-protein interactions amongst identified transcription factors mediating PTSD hyper-responsivity.
  • FIG.5C is a graph showing the overlap of transcription factors (dashed) and their targets (white) identified in the study with significantly differentially expressed genes in other PTSD studies.
  • FIG.5D is a set of Manhattan plots of significantly interactive genes in the study compared to Manhattan plot of imputed expression from PTSD genome-wide association studies (GWAS) indicating spatial orientation of significantly interactive genes.
  • FIG.6 is a set of photomicrographs showing the immunostaining of hiPSC- derived NGN2-neurons. Immunostaining of Hoechst, GFP, MAP2, and NESTIN across all participants.
  • FIGs.7A-7B are a set of graphs showing adjustment of Batch effects.
  • FIG.7A is a set of graphs showing VariancePartition (left), primary component analysis (PCA; middle), and an example gene after batch correction (right). This analysis on uncorrected data indicate a large batch effect.
  • FIG.7B is a set of graphs showing VariancePartition (left), PCA (middle), and an example gene after batch correction (right).
  • FIGs.8A-8B are a set of graphs showing a developmental specificity analysis.
  • FIG.8A is a graph showing pair-wise correlation between NGN2s from the study with cell types across 16 independent studies
  • FIG.8B is a graph showing the PCA analysis of cell types within all 16 studies with the NGN2 neurons.
  • FIGs.9A-9C are a set of graphs showing a neuronal fate specificity analysis.
  • FIG.9A is a heat map showing the expression of hallmark pan-neuronal and neuronal subtype specific genes in NGN2 neurons and PBMCs.
  • FIG.9B is a graph showing the average log2CPM expression of vesicular glutamate transporter 1 (VGLUT1) and vesicular glutamate transporter 2 (VGLUT2) in NGN2 neurons and PBMCs.
  • FIG.9C is a graph showing the expression of GR and mineralocorticoid (MR) in NGN2 neurons and PBMCs.
  • FIGs.10A-10D are a set of graphs showing the comparison of PBMC batches.
  • FIG.10A is a graph showing pair-wise correlations between PBMC batches.
  • FIG.10B is a graph showing the transcriptome-wide correlation between batches at the 50 nM DEX dose.
  • FIG.10C is a graph showing the pair-wise correlations between PBMC batches and Breen, M. S. et al. Translational Psychiatry 9, 201, doi:10.1038/s41398-019-0539-x (2019).
  • FIG. 10D is a graph showing the transcriptome-wide correlation between the study and Breen et al. 2019 at the 50 nM DEX dose.
  • FIGs.11A-11B are a set of graphs showing a comparison of NGN2 batches.
  • FIG. 11A is a set of graphs showing HCort-responsive DEGs across independent batches. Transcriptome-wide concordance is plotted between dosages for each batch.
  • FIG.11B is a graph showing the quantification of cell number with HCort treatment showing no significant cell density between doses.
  • FIGs.12A-12B are a set of graphs showing a weighted gene co-expression network analysis of PBMCs (FIG.12A) and NGN2 neurons (FIG.12B).
  • Hierarchical gene cluster tree, module structure, and gene-treatment are denoted by gray bands. The first band underneath the tree indicates the detected modules and subsequent bands indicate treatment correlation, where lighter gray indicates a strong relationship and darker gray indicates a strong negative relationship.
  • FIGs.13A-13B are a set of graphs showing analysis of unsigned modules.
  • FIG.13B is a set of network visualization of protein- protein interactions within unsigned modules indicating clusters and network hubs.
  • FIGs.14A-14B are a set of graphs showing PTSD-positive-specific responses to DEX in PBMCs.
  • FIG.14A is a graph showing genes that differ in their response to DEX in PTSD-positive donors compared to PTSD-negative donors, here termed “differential response genes (DRGs),” at a false discovery rate (FDR) threshold of 20% (non-significant).
  • FIG.14B is a set of graphs showing unsupervised clustering of nominally significant PTSD DRGs.
  • FIG.15 is a graph (left) and Venn diagram (right), respectively, showing the concordance of PBMC signature with NGN2 signature. Pair-wise correlations between transcriptome-wide signatures of PBMC and NGN2 batches are shown.
  • the term “about” means within 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.
  • “activity” refers to form(s) of a gene or respectively encoded protein which retains a biological activity of the native or naturally-occurring gene or polypeptide, respectively.
  • the term “administering,” or a grammatical derivative thereof, as described herein, refers to the delivery of an agent, e.g., an agent that modifies the glucocorticoid response and/or a psychedelic agent to an individual in need thereof. Any suitable method of administration can be selected by one of skill in the art, in view of this disclosure.
  • an agent is administered via a parenteral route. In some embodiments, an agent is administered via a non-parenteral route.
  • biological sample or “sample” is meant a fluid or solid sample from an individual. Biological samples may include cells (e.g., neurons (e.g., glutamatergic neurons), blood cells (e.g., peripheral blood mononuclear cells), human induced pluripotent cells (hIPSc); nucleic acid, protein, or membrane extracts of cells; or blood or biological fluids including (e.g., plasma, serum, saliva, urine, bile).
  • cells e.g., neurons (e.g., glutamatergic neurons), blood cells (e.g., peripheral blood mononuclear cells), human induced pluripotent cells (hIPSc); nucleic acid, protein, or membrane extracts of cells; or blood or biological fluids including (e.g., plasma, serum, saliva, urine, bile).
  • Solid biological samples include samples taken from feces, the rectum, central nervous system, bone, breast tissue, renal tissue, the uterine cervix, the endometrium, the head or neck, the gallbladder, parotid tissue, the prostate, the brain, the pituitary gland, kidney tissue, muscle, the esophagus, the stomach, the small intestine, the colon, the liver, the spleen, the pancreas, thyroid tissue, heart tissue, lung tissue, the bladder, adipose tissue, lymph node tissue, the uterus, ovarian tissue, adrenal tissue, testis tissue, the tonsils, and the thymus.
  • Fluid biological samples include samples taken from the blood, serum, plasma, pancreatic fluid, CSF, semen, prostate fluid, seminal fluid, urine, saliva, sputum, mucus, bone marrow, lymph, and tears.
  • the biological sample is a blood, plasma, or serum sample.
  • the biological sample includes blood cells (e.g., peripheral blood mononuclear cells), neurons (e.g., glutamatergic neurons), fibroblasts, or cells later derived into hiPSC. Samples may be obtained by standard methods including, e.g., skin puncture and surgical biopsy.
  • a biological sample includes one or more cells, which are processed to produce a test cell.
  • control means any useful reference used to compare the expression and/or activity of the one or more genes of the glucocorticoid response.
  • the baseline can be any sample, standard, standard curve, or level that is used for comparison purposes.
  • the baseline can be a normal reference sample or a reference standard or level.
  • a “suitable control” can be, for example, a control, e.g., a predetermined negative control value such as a “normal control” or a prior sample taken from the same individual; a sample from a normal healthy individual, a sample from an individual not having PTSD; or a sample from an individual that has been treated for PTSD.
  • reference standard or level is meant a value or number derived from a reference sample.
  • a “normal control value” is a pre-determined value indicative of non-disease state, e.g., a value expected in a healthy control individual. Typically, a normal control value is expressed as a range (“between X and Y”), a high threshold (“no higher than X”), or a low threshold (“no lower than X”).
  • An individual having a measured value within the normal control value for a particular assay is typically referred to as “within normal limits” for that assay.
  • a normal reference standard or level can be a value or number derived from a normal individual not having PTSD; or an individual that has been treated for PTSD.
  • the reference sample, standard, or level is matched to the sample individual sample by at least one of the following criteria: age, weight, sex, disease stage, and overall health.
  • a “suitable control” refers to the expression and/or activity levels of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response against which the expression and/or activity levels of the respective genes are compared, e.g., to make a diagnostic, predictive, prognostic, and/or therapeutic determination.
  • a suitable control includes substantially no test agent administered to an individual.
  • the word “comprise,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated word or group of words but not the exclusion of any other word or group of words.
  • the term “detection” includes any means of detecting, including direct and indirect detection.
  • determining the level of a nucleic acid is meant the detection of a nucleic acid (e.g., mRNA) by methods known in the art.
  • Methods to measure mRNA levels generally include, but are not limited to, northern blotting, nuclease protection assays (NPA), in situ hybridization (ISH), reverse transcription-polymerase chain reaction (RT-PCR), and RNA sequencing (RNA-Seq).
  • NPA nuclease protection assays
  • ISH in situ hybridization
  • RT-PCR reverse transcription-polymerase chain reaction
  • RNA-Seq RNA sequencing
  • Methods to measure protein levels generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, florescent polarization, phosphorescence, immunohistochemical analyses, matrix-associated laser desorption/ionization time of light (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS), and flow cytometry, as well as assays based on a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners.
  • MALDI-TOF matrix-associated laser desorption/ionization time of light
  • LC liquid chromatography
  • FACS fluorescence activated cell sorting
  • flow cytometry as well assays based on a property of a
  • diagnosis refers to the identification or classification of a genetic, molecular, or pathological state, disease, or condition (e.g., PTSD).
  • diagnosis may refer to identification of an individual with PTSD.
  • effective amount refers to a quantity sufficient to, when administered to an individual, including human, effect beneficial or desired results (e.g., alleviate one or more symptoms of PTSD), which may include clinical results.
  • an effective amount of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents described herein may alleviate one or more symptoms of PTSD as compared to the alleviation of said symptom without administration of the agent of interest.
  • agents described herein e.g., agents that modify the glucocorticoid response or psychedelic agents
  • An “effective amount,” “therapeutically effective amount,” and the like, of an agent, such as a glucocorticoid receptor antagonist or a psychedelic agent also include an amount that results in a beneficial or desired result in an individual as compared to a control.
  • glucocorticoid receptor antagonist is a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of a glucocorticoid receptor with either one or more of its binding partners.
  • glucocorticoid receptor antagonist is a molecule that inhibits the binding of a glucocorticoid receptor to its binding partners.
  • glucocorticoid receptor antagonists include small molecule antagonists, polynucleotide antagonists, antibodies and antigen-binding fragments thereof, fusion proteins, oligopeptides, and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of a glucocorticoid receptor with one or more of its binding partners.
  • identifying an individual or “identifies an individual,” as used herein, refers to using the information or data generated related to the expression and/or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response to identify or select an individual as likely to benefit or less likely to benefit from a therapy including one or more psychedelic agents and/or agents that modify the glucocorticoid response, including a glucocorticoid receptor antagonist.
  • the information or data used or generated may by be in any form, written, oral, or electronic.
  • using the information or data generated includes communicating, presenting, reporting, storing, sending, transferring, supplying, transmitting, dispensing, or combinations thereof.
  • communicating, presenting, reporting, storing, sending, transferring, supplying, transmitting, dispensing, or combinations thereof are performed by a computing device, analyzer unit, or combination thereof.
  • the information or data includes a comparison of the expression and/or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response to a reference level (e.g., a level from a suitable control).
  • the information or data includes an indication that the expression and/or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response are elevated relative to a suitable control (e.g., a control with substantially no test agent).
  • a suitable control e.g., a control with substantially no test agent.
  • the information or data includes an indication that the individual has or does not have an elevated risk for PTSD.
  • the terms “induced pluripotent stem cell,” “iPS cell,” and “iPSC” refer to a pluripotent stem cell that can be derived directly from a differentiated somatic cell.
  • Human iPS cells can be generated by introducing specific sets of reprogramming factors into a non-pluripotent cell (e.g., fibroblasts) that can include, for example, Oct-3/4, Sox family, Klf family, Myc family, Nanog, LIN28, and Glis1 genes.
  • Human iPS cells can also be generated, for example, by the use of miRNAs, small molecules that mimic the actions of transcription factors, or lineage specifiers.
  • Human iPS cells are characterized by their ability to differentiate into any cell of the three vertebrate germ layers, e.g., the endoderm, the ectoderm, or the mesoderm.
  • Human iPS cells are also charactered by their ability to propagate indefinitely under suitable in vitro culture conditions. See, for example, Takahashi and Yamanaka, Cell 126:663 (2006).
  • a cell obtained from a biological sample is dedifferentiated to produce an iPSC.
  • the iPSC is then differentiated to produce a test cell.
  • the iPSC may be differentiated into an induced neuron or an induced peripheral blood mononuclear cell.
  • a cell obtained from a biological sample is processed by automated reprogramming.
  • level is meant a level of a genes expression or activity as compared to a reference.
  • the reference can be any useful reference, as defined herein.
  • a “decreased level” or an “increased level” of a gene is meant a decrease or increase in gene expression or activity, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by less than about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less; or an increase by more than about 1.2-fold, about 1.4-fold, about 1.5
  • level of expression or “expression level” in general are used interchangeably and generally refer to the amount of a biomarker (e.g., one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response) in a biological sample). “Expression” generally refers to the process by which information (e.g., gene-encoded and/or epigenetic information) is converted into the structures present and operating in a cell.
  • expression may refer to transcription into a polynucleotide translation into a polypeptide, or even polynucleotide and/or polypeptide modifications (e.g., posttranslational modification of a polypeptide). Fragments of the transcribed polynucleotide, the translated polypeptide, or polynucleotide and/or polypeptide modifications (e.g., post-translational modifications of a polypeptide) shall also be regarded as expressed whether they originate from a transcript generated by alternative splicing or a degraded transcript, or from post-translational processing of a polypeptide, e.g., by proteolysis.
  • modified refers to an observable difference in the level of a marker, such as the expression and/or activity of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) gene(s), in a sample (e.g., a biological sample from an individual e.g., an individual suspected of being at risk for PTSD or diagnosed with PTSD), as determined using techniques and methods known in the art for the measurement of the marker.
  • a sample e.g., a biological sample from an individual e.g., an individual suspected of being at risk for PTSD or diagnosed with PTSD
  • a marker level that is changed in an individual may result in a difference of at least 1% (e.g., at least 5%, 10%, 25%, 50%, or 100% or at least 2.5-fold, 3-fold, 4-fold, 5-fold, 6- fold, 7-fold or more) compared to a reference level, e.g., a level from a suitable control.
  • the change is an increased level of the expression or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response in a biological sample from an individual.
  • the change is a decreased level of the expression or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response in a biological sample from an individual.
  • the one or more e.g., two, three, four, five, ten, fifteen, or twenty or more
  • post-traumatic stress disorder and “PTSD” are defined for the purposes of the present disclosure as a potentially debilitating anxiety disorder triggered by exposure to trauma or a traumatic experience (e.g., a catastrophic or threatening event, e.g., a natural disaster, a wartime situation, an accident, domestic abuse, or a violent crime), such as an interpersonal event associated with actual or threatening death or severe death, such as, for example, physical or sexual assault; exposure to disaster or accidents; combat; or witnessing a traumatic event.
  • trauma or a traumatic experience e.g., a catastrophic or threatening event, e.g., a natural disaster, a wartime situation, an accident, domestic abuse, or a violent crime
  • an interpersonal event associated with actual or threatening death or severe death such as, for example, physical or sexual assault
  • exposure to disaster or accidents combat
  • combat or witnessing a traumatic event.
  • PTSD symptoms There are three main clusters of PTSD symptoms: firstly, those related to re-experiencing the event; secondly, those related to avoidance and arousal; and thirdly, the distress and impairment caused by the first two symptom clusters.
  • the terms “individual,” “subject,” and “patient” are used interchangeably and are meant as a human .
  • An individual to be treated with a pharmaceutical composition described herein may be one who has been diagnosed by a medical practitioner as having PTSD or one at risk for developing PTSD.
  • the terms “treat,” “treatment,” “treating,” and the like are used herein to generally mean obtaining a desired pharmacological and/or physiological effect.
  • treatment covers any treatment of PTSD in a human, and includes: (a) inhibiting the disorder, i.e., preventing the disorder from increasing in severity or scope; (b) relieving the disorder, i.e., causing partial or complete amelioration of the disorder; or (c) preventing relapse of the disorder, i.e., preventing the disorder from returning to an active state following previous successful treatment of symptoms of the disorder or treatment of the disorder.
  • the term “reference level” refers to the level as determined in a suitable control as further described herein, e.g., a level from a prior biological sample taken from the same individual, a biological sample from a healthy individual not having a risk for PTSD, a biological sample from an individual not having one or more symptoms associated with PTSD, or a biological sample from an individual that is diagnosed with PTSD that has been treated for PTSD.
  • Post-Traumatic Stress Disorder [0089] The methods described herein can be used to diagnose, prognose, and treat post- traumatic stress disorder (PTSD). PTSD is a dominant and highly debilitating psychiatric disorder that is notoriously difficult to treat.
  • PTSD can be characterized by intrusive recall, emotional numbness, and insomnia and is associated with functional deficiencies, physical health problems, and mental health comorbidities such as depression, with a six-fold increased risk of suicide.
  • PTSD can result from a catastrophic or threatening event, e.g., a natural disaster, a wartime situation, an accident, domestic abuse, or a violent crime. Symptoms normally develop over the course of three months, but may emerge years after the initial trauma.
  • DSM-IV-TR ® describes post-traumatic stress disorder (PTSD) as developing characteristic symptoms after exposure to extreme traumatic stressors, including direct personal experience of events associated with actual or threatening death or severe death.
  • a person with PTSD can be a witness of an event that includes the death, injury, or threat to physical integrity of another person. Human reactions to events include, but are not limited to, intense fear, lethargy, or fear.
  • a person with PTSD can have a persistent memory of an event, including images, thoughts, or perceptions, or can have a recurring painful dream of the event.
  • Genes of the Glucocorticoid Response [0091] Applicants have discovered that the mRNA expression levels and/or activity of certain genes can be utilized to diagnose, prognose, and treat PTSD, as well as to select individuals who would benefit from a treatment that modifies the glucocorticoid response.
  • genes can also be used for screening compounds that reduce the risk of an individual developing PTSD, reduce the risk of an individual developing one or more symptoms of PTSD, and/or alleviate one or more symptoms of PTSD.
  • biomarkers e.g., one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response (e.g., genes of Table 1 and/or Table 2) that identify individuals at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD.
  • the differential expression and/or activity levels of genes of the glucocorticoid response e.g., genes of PTSD-dependent glucocorticoid response gene signature
  • suitable controls e.g., healthy controls.
  • the methods described herein are useful for treating or diagnosing PTSD.
  • Diagnostic and Classification Methods [0093]
  • the present disclosure features methods to diagnose PTSD. Methods of the present disclosure may be used alone or as a companion diagnostics with other diagnostic or therapeutic approaches, as an early molecular screen to distinguish PTSD.
  • alterations in the expression level and/or activity of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response exemplified herein (e.g., one or more genes (e.g., genes of Table 1 and/or Table 2) in a biological sample (e.g., blood cell (e.g., peripheral blood mononuclear cell), neuron (e.g., glutamatergic neuron), fibroblast, or hiPSC) from the individual suspected of being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD as compared to a suitable control (e.g., a normal reference such as a control with substantially no test agent) can be used to diagnose PTSD from diseases or disorders with similar symptoms, thereby allowing individual classification.
  • a biological sample e.g., blood cell (e.g., peripheral blood mononuclear cell), neuron (e.g., glutamatergic
  • the method includes processing a cell obtained from the biological sample to produce a test cell.
  • the method further includes contacting the biological sample or cells with a glucocorticoid (e.g., dexamethasone or hydrocortisone) prior to detecting the expression level and/or activity of one or more genes of the glucocorticoid response.
  • a glucocorticoid e.g., dexamethasone or hydrocortisone
  • the method includes identifying an individual at risk for PTSD or diagnosed with PTSD by obtaining a biological sample from the individual suspected of being at risk for PTSD or diagnosed with PTSD; processing a cell obtained from the biological sample to produce a test cell; contacting the test cell with a glucocorticoid (e.g., dexamethasone or hydrocortisone) to produce a glucocorticoid-induced response; detecting the expression and/or activity of one or more (e.g., (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid-induced response (e.g., one or more genes of Table 1 and/or Table 2); and identifying the individual as at risk for PTSD or diagnosing the individual with PTSD if the expression and/or activity of the one or more genes of the glucocorticoid-induced response are modified relative to a suitable control (e.g.,
  • Methods of the present disclosure can be used to diagnose, prognose, or classify an individual, for example, an increase in the expression and/or activity (e.g., an increase by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 300%, 400%, 500%, or more, or an increase by more than 1.2-fold, 1.4-fold, 1.5-fold, 1.8-fold, 2.0-fold, 3.0-fold, 3.5-fold, 4.5-fold, 5.0-fold, 10- fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, or more, as compared to a reference) of the biomarkers (e.g., one or more genes of Table 1 and/or Table 2) may identify an individual as being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD and
  • a decrease in the level e.g., a decrease by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 300%, 400%, 500%, or more; or a decrease by less than 0.01-fold, 0.02-fold, 0.1-fold, 0.3-fold, 0.5-fold, 0.8-fold, or less, as compared to a reference) of the biomarkers (e.g., one or more genes of Table 1 and/or Table 2) may identify an individual as being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD and/or may benefit from one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist) and/or psych
  • the present disclosure further features methods for predicting response to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist) in cells from individuals at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD, before or after administration of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist).
  • agents that modify the glucocorticoid response e.g., a glucocorticoid receptor antagonist
  • the method includes screening compounds that reduce the risk of an individual developing PTSD, reduce the risk of an individual developing one or more symptoms of PTSD, and/or alleviate one or more symptoms of PTSD in an individual by obtaining a biological sample from the individual at risk for PTSD or suffering from PTSD; processing a cell obtained from the biological sample to produce a test cell; contacting the test cell with a glucocorticoid to produce a glucocorticoid-induced response; contacting the test cell with one or more test agents; detecting the expression and/or activity of one or more genes of the glucocorticoid-induced response (e.g., genes of Table 1 and/or Table 2); and if the one or more test agents modifies the expression and/or activity of one or more genes of the glucocorticoid-induced response (e.g., genes of Table 1 and/or Table 2) compared to a suitable control (e.g., a control with substantially no test agent), identifying the test agent as
  • these methods may be carried out generally as described above or as known in the art with respect to sample collection and assay format.
  • these methods may be carried out by obtaining cells from individuals at risk for PTSD or suffering from PTSD; contacting the cells with one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) test agents; detecting the expression and/or activity of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response (e.g., genes of Table 1 and/or Table 2) in the sample and/or determining if the test agent reduces the transcriptional profile of the PTSD- dependent glucocorticoid response gene signature; and making a prediction about whether a test agent may reduce the risk of an individual developing PTSD, reduce the risk of an individual developing one or more symptoms of PTSD, and/or alleviate one or more symptoms of PTSD in an individual.
  • one or more e.g., two, three,
  • the method also can be used to predict whether an individual, who has been diagnosed with PTSD, will respond positively to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist).
  • the method includes processing a cell obtained from the biological sample to produce a test cell.
  • the method further includes contacting the biological sample or cells with a glucocorticoid (e.g., dexamethasone or hydrocortisone) prior to detecting the expression level and/or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response.
  • a glucocorticoid e.g., dexamethasone or hydrocortisone
  • a prediction of a positive response refers to a case where the PTSD symptoms will be alleviated as a result of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist).
  • agents that modify the glucocorticoid response e.g., a glucocorticoid receptor antagonist
  • the transcriptional profile of the PTSD-dependent glucocorticoid response gene signature can be determined relative to a control value.
  • a control value can be a range or average value from a normal individual or a population of normal individuals; a value from a sample from an individual or population of individuals who have undergone one or more agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist) and have reduced symptoms following therapy; or a value from the same individual before the individual was diagnosed or before the individual started treatment.
  • agents that modify the glucocorticoid response e.g., a glucocorticoid receptor antagonist
  • Methods of the present disclosure can be used to predict whether an individual will be responsive to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist), for example, an increase in the level (e.g., an increase by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 300%, 400%, 500%, or more, or an increase by more than 1.2-fold, 1.4-fold, 1.5-fold, 1.8- fold, 2.0-fold, 3.0-fold, 3.5-fold, 4.5-fold, 5.0-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 100-fold, 1000-fold, or more, as compared to a reference) of
  • a decrease in the level may indicate a positive response to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist).
  • Methods of the present disclosure can be used to predict an individual’s response to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist) and classify the individual as a “responder,” e.g., an individual with a glucocorticoid response gene signature indicative of a positive response to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist), or a “non-responder,” e.g., an individual with a glucocorticoid response gene signature indicative of a poor response to one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocortic
  • the prediction can be made prior to administration of a first agent that modifies the glucocorticoid response (e.g., a glucocorticoid receptor antagonist). Alternatively, the prediction can be made after administration of the first agent that modifies the glucocorticoid response (e.g., a glucocorticoid receptor antagonist), or after administration of a first agent that modifies the glucocorticoid response (e.g., a glucocorticoid receptor antagonist) but before a second agent that modifies the glucocorticoid response.
  • a first agent that modifies the glucocorticoid response e.g., a glucocorticoid receptor antagonist
  • the prediction can be made at any time during the course of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) agents that modify the glucocorticoid response (e.g., a glucocorticoid receptor antagonist).
  • agents that modify the glucocorticoid response e.g., a glucocorticoid receptor antagonist.
  • the methods described herein can also be used to monitor PTSD status (e.g., progression or regression) during therapy or to optimize dosage of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) therapeutic agents for an individual.
  • alterations e.g., an increase or a decrease as compared to either the positive reference sample or the level diagnostic for PTSD
  • the levels of the glucocorticoid response gene signature may be measured repeatedly as a method of not only diagnosing disorder, but also monitoring the treatment, prevention, or management of the disorder.
  • individual samples may be compared to reference samples taken early in the diagnosis of the disorder. Such monitoring may be useful, for example, in assessing the efficacy of a particular therapeutic agent (e.g., a glucocorticoid receptor antagonist) in an individual, determining dosages, or in assessing disease progression or status.
  • a particular therapeutic agent e.g., a glucocorticoid receptor antagonist
  • the expression and/or activity of any of the genes described herein, or any combination thereof can be monitored in an individual, and as the expression levels or activities increase or decrease, relative to control, the dosage or administration of therapeutic agents may be adjusted.
  • the methods can also be used to determine the proper dosage (e.g., the therapeutically effective amount) of a therapeutic agent for the individual, the proper type of therapeutic agent, or whether a therapy should be administered.
  • Methods of Treatment [00110] The present disclosure also features a method of treating an individual diagnosed with PTSD including administering to the individual diagnosed with PTSD a therapeutically effective amount of a psychedelic agent and/or a glucocorticoid receptor antagonist.
  • the present disclosure also features a method for treatment of PTSD in an individual by obtaining a biological sample (e.g., blood cell (e.g., peripheral blood mononuclear cell), neuron (e.g., glutamatergic neuron), fibroblast, or hiPSC) from the individual suspected of being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD; detecting the expression and/or activity of one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response (e.g., one or more genes of Table 1 and/or Table 2); identifying an individual at risk for PTSD or diagnosed with PTSD when the expression and/or activity of the one or more (e.g., two, three, four, five, ten, fifteen, or twenty or more) genes of the glucocorticoid response are modified relative to a suitable control (e.g., a control with substantially no test agent);
  • the method includes processing a cell obtained from the biological sample to produce a test cell.
  • the method further includes contacting the biological sample or cells with a glucocorticoid (e.g., dexamethasone or hydrocortisone) prior to detecting the expression level and/or activity of one or more genes of the glucocorticoid response.
  • a glucocorticoid e.g., dexamethasone or hydrocortisone
  • the method includes treating an individual at risk for PTSD, suffering from one or more symptoms associated with PTSD, and/or diagnosed with PTSD by obtaining a biological sample from the individual suspected of being at risk for PTSD, suffering from one or more symptoms associated with PTSD, or diagnosed with PTSD; processing a cell obtained from the biological sample to produce a test cell; contacting the test cell with a glucocorticoid (e.g., dexamethasone or hydrocortisone) to produce a glucocorticoid-induced response; detecting the expression and/or activity of one or more genes of the glucocorticoid-induced response (e.g., one or more genes of Table 1 and/or Table 2); identifying the individual as at risk for PTSD or diagnosing the individual with PTSD when the expression and/or activity of the one or more genes of the glucocorticoid- induced response are modified relative to a suitable control (e.g., a control with substantially no
  • the method includes treating an individual at risk for developing PTSD, diagnosed with PTSD, or experiencing one or more symptoms associated with PTSD by administering to the individual one more psychedelic agents and/or agents that modify the glucocorticoid-induced response (e.g., a glucocorticoid receptor antagonist).
  • the methods of the disclosure