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Jagadeesh, K. A.

Publications and source records attributed to Jagadeesh, K. A..

10 recordsLinked to original sources

Polygenic enrichment distinguishes disease associations of individual cells in single-cell RNA-seq data

Gene expression at the individual cell-level resolution, as quantified by single-cell RNA-sequencing (scRNA-seq), can provide unique insights into the pathology and cellular origin of diseases and complex traits. Here, we introduce single-cell Disease Relevance Score (scDRS), an approach that links scRNA-seq with polygenic risk of disease at individual cell resolution without the need for annotation of individual cells to cell types; scDRS identifies individual cells that show excess expression levels for genes in a disease-specific gene set constructed from GWAS data. We determined via simulations that scDRS is well-calibrated and powerful in identifying individual cells associated to disease. We applied scDRS to GWAS data from 74 diseases and complex traits (average N =346K) in conjunction with 16 scRNA-seq data sets spanning 1.3 million cells from 31 tissues and organs. At the cell type level, scDRS broadly recapitulated known links between classical cell types and disease, and also produced novel biologically plausible findings. At the individual cell level, scDRS identified subpopulations of disease-associated cells that are not captured by existing cell type labels, including subpopulations of CD4+ T cells associated with inflammatory bowel disease, partially characterized by their effector-like states; subpopulations of hippocampal CA1 pyramidal neurons associated with schizophrenia, partially characterized by their spatial location at the proximal part of the hippocampal CA1 region; and subpopulations of hepatocytes associated with triglyceride levels, partially characterized by their higher ploidy levels. At the gene level, we determined that genes whose expression across individual cells was correlated with the scDRS score (thus reflecting co-expression with GWAS disease genes) were strongly enriched for gold-standard drug target and Mendelian disease genes.

bioinformatics↗

Identifying disease-critical cell types and cellular processes across the human body by integration of single-cell profiles and human genetics

Genome-wide association studies (GWAS) provide a powerful means to identify loci and genes contributing to disease, but in many cases the related cell types/states through which genes confer disease risk remain unknown. Deciphering such relationships is important for identifying pathogenic processes and developing therapeutics. Here, we introduce sc-linker, a framework for integrating single-cell RNA-seq (scRNA-seq), epigenomic maps and GWAS summary statistics to infer the underlying cell types and processes by which genetic variants influence disease. We analyzed 1.6 million scRNA-seq profiles from 209 individuals spanning 11 tissue types and 6 disease conditions, and constructed gene programs capturing cell types, disease progression, and cellular processes both within and across cell types. We evaluated these gene programs for disease enrichment by transforming them to SNP annotations with tissue-specific epigenomic maps and computing enrichment scores across 60 diseases and complex traits (average N=297K). Cell type, disease progression, and cellular process programs captured distinct heritability signals even within the same cell type, as we show in multiple complex diseases that affect the brain (Alzheimers disease, multiple sclerosis), colon (ulcerative colitis) and lung (asthma, idiopathic pulmonary fibrosis, severe COVID-19). The inferred disease enrichments recapitulated known biology and highlighted novel cell-disease relationships, including GABAergic neurons in major depressive disorder (MDD), a disease progression M cell program in ulcerative colitis, and a disease-specific complement cascade process in multiple sclerosis. In autoimmune disease, both healthy and disease progression immune cell type programs were associated, whereas for epithelial cells, disease progression programs were most prominent, perhaps suggesting a role in disease progression over initiation. Our framework provides a powerful approach for identifying the cell types and cellular processes by which genetic variants influence disease.

genomics↗

A single-cell and spatial atlas of autopsy tissues reveals pathology and cellular targets of SARS-CoV-2

The SARS-CoV-2 pandemic has caused over 1 million deaths globally, mostly due to acute lung injury and acute respiratory distress syndrome, or direct complications resulting in multiple-organ failures. Little is known about the host tissue immune and cellular responses associated with COVID-19 infection, symptoms, and lethality. To address this, we collected tissues from 11 organs during the clinical autopsy of 17 individuals who succumbed to COVID-19, resulting in a tissue bank of approximately 420 specimens. We generated comprehensive cellular maps capturing COVID-19 biology related to patients demise through single-cell and single-nucleus RNA-Seq of lung, kidney, liver and heart tissues, and further contextualized our findings through spatial RNA profiling of distinct lung regions. We developed a computational framework that incorporates removal of ambient RNA and automated cell type annotation to facilitate comparison with other healthy and diseased tissue atlases. In the lung, we uncovered significantly altered transcriptional programs within the epithelial, immune, and stromal compartments and cell intrinsic changes in multiple cell types relative to lung tissue from healthy controls. We observed evidence of: alveolar type 2 (AT2) differentiation replacing depleted alveolar type 1 (AT1) lung epithelial cells, as previously seen in fibrosis; a concomitant increase in myofibroblasts reflective of defective tissue repair; and, putative TP63+ intrapulmonary basal-like progenitor (IPBLP) cells, similar to cells identified in H1N1 influenza, that may serve as an emergency cellular reserve for severely damaged alveoli. Together, these findings suggest the activation and failure of multiple avenues for regeneration of the epithelium in these terminal lungs. SARS-CoV-2 RNA reads were enriched in lung mononuclear phagocytic cells and endothelial cells, and these cells expressed distinct host response transcriptional programs. We corroborated the compositional and transcriptional changes in lung tissue through spatial analysis of RNA profiles in situ and distinguished unique tissue host responses between regions with and without viral RNA, and in COVID-19 donor tissues relative to healthy lung. Finally, we analyzed genetic regions implicated in COVID-19 GWAS with transcriptomic data to implicate specific cell types and genes associated with disease severity. Overall, our COVID-19 cell atlas is a foundational dataset to better understand the biological impact of SARS-CoV-2 infection across the human body and empowers the identification of new therapeutic interventions and prevention strategies.

genomics↗

Single-nucleus and spatial transcriptomics of archival pancreatic cancer reveals multi-compartment reprogramming after neoadjuvant treatment

Pancreatic ductal adenocarcinoma (PDAC) remains a treatment-refractory disease. Characterizing PDAC by mRNA profiling remains particularly challenging. Previously identified bulk expression subtypes were influenced by contaminating stroma and have not yet informed clinical management, whereas single cell RNA-seq (scRNA-seq) of fresh tumors under-represented key cell types. Here, we developed a robust single-nucleus RNA-seq (snRNA-seq) technique for frozen archival PDAC specimens and used it to study both untreated tumors and those that received neoadjuvant chemotherapy and radiotherapy (CRT). Gene expression programs learned across untreated malignant cell and fibroblast profiles uncovered a clinically relevant molecular taxonomy with improved prognostic stratification compared to prior classifications. Moreover, in the increasingly-adopted neoadjuvant treatment context, there was a depletion of classical-like phenotypes in malignant cells in favor of basal-like phenotypes associated with TNF-NFkB and interferon signaling as well as the presence of novel acinar and neuroendocrine classical-like states, which may be more resilient to cytotoxic treatment. Spatially-resolved transcriptomics revealed an association between malignant cells expressing these basal-like programs and higher immune infiltration with increased lymphocytic content, whereas those exhibiting classical-like programs were linked to sparser macrophage-predominant microniches, perhaps pointing to susceptibility to distinct therapeutic strategies. Our refined molecular taxonomy and spatial resolution can help advance precision oncology in PDAC through informative stratification in clinical trials and insights into differential therapeutic targeting leveraging the immune system.

genomics↗

Innocent until proven guilty: Privacy-preserving search over a central CODIS criminal database from the field

The presumption of innocence (i.e., the principle that one is considered innocent until proven guilty) is a cornerstone of the criminal justice system in many countries, including the United States. DNA analysis is an important tool for criminal investigations1. In the U.S. alone, it has already aided in over half a million investigations using the Combined DNA Index System (CODIS) and associated DNA databases2. CODIS includes DNA profiles of crime scene forensic samples, convicted offenders, missing persons and more. The CODIS framework is currently used by over 50 other countries3 including much of Europe, Canada, China and more. During investigations, DNA samples can be collected from multiple individuals who may have had access to, or were found near a crime scene, in the hope of finding a single criminal match4. Controversially, CODIS samples are sometimes retained from adults and juveniles despite not yielding any database match4-6. Here we introduce a cryptographic algorithm that finds any and all matches of a persons DNA profile against a CODIS database without revealing anything about the persons profile to the database provider. With our protocol, matches are immediately identified as before; however, individuals who do not match anything in the database retain their full privacy. Our novel algorithm runs in 40 seconds on a CODIS database of 1,000,000 entries, enabling its use to privately screen potentially-innocent suspects even in the field.

genomics↗

AMELIE 2 speeds up Mendelian diagnosis by matching patient phenotype & genotype to primary literature

The diagnosis of Mendelian disorders requires labor-intensive literature research. Trained clinicians can spend hours looking for the right publication/s supporting a single gene that best explains a patients disease. AMELIE (Automatic Mendelian Literature Evaluation) greatly accelerates this process. AMELIE parses all 29 million PubMed abstracts, downloads and further parses hundreds of thousands of full text articles in search of information supporting the causality and associated phenotypes of any published genetic variant. AMELIE then prioritizes patient candidate variants for their likelihood of explaining any patients given set of phenotypes. Diagnosis of singleton patients (without relatives exomes) is the most time-consuming scenario. AMELIE ranked the causative gene at the very top in 2/3 of 215 diagnosed singleton Mendelian patients. Evaluating only the top 11 AMELIE scored genes of 127 (median) candidate genes per patient results in rapid diagnosis for 90+% of cases. AMELIE-based evaluation of all cases is 3-19x more efficient than hand-curated database-based approaches. We replicate these results on a cohort of clinical cases from Stanford Childrens Health and the Manton Center for Orphan Disease Research. An analysis web portal with our most recent update, programmatic interface and code will be available at AMELIE.stanford.edu. A pilot run of the web portal has already served many thousands of job submissions from dozens of countries.

genetics↗

AVADA Enables Automated Genetic Variant Curation Directly from the Full Text Literature

PurposeThe primary literature on human genetic diseases includes descriptions of pathogenic variants that are essential for clinical diagnosis. Variant databases such as ClinVar and HGMD collect pathogenic variants by manual curation. We aimed to automatically construct a freely accessible database of pathogenic variants directly from full-text articles about genetic disease.\n\nMethodsAVADA (Automatically curated VAriant DAtabase) is a novel machine learning tool that uses natural language processing to automatically identify pathogenic variants and genes in full text of primary literature and converts them to genomic coordinates for rapid downstream use.\n\nResultsAVADA automatically curated almost 60% of pathogenic variants deposited in HGMD, a 4.4-fold improvement over the current state of the art in automated variant extraction. AVADA also contains more than 60,000 pathogenic variants that are in HGMD, but not in ClinVar. In a cohort of 245 diagnosed patients, AVADA correctly annotated 38 previously described diagnostic variants, compared to 43 using HGMD, 20 using ClinVar and only 13 (wholly subsumed by AVADA and ClinVars) using the best automated abstracts-only based approach.\n\nConclusionAVADA is the first machine learning tool that automatically curates a variants database directly from full text literature. AVADA is available upon publication at http://bejerano.stanford.edu/AVADA.

genetics↗

S-CAP extends clinical-grade pathogenicity prediction to genetic variants that affect RNA splicing

There are over 15,000 known variants that cause human inherited disease by disrupting RNA splicing. While several in silico methods such as CADD, EIGEN and LINSIGHT are commonly used to predict the pathogenicity of noncoding variants, we introduce S-CAP, a tool developed specially for splicing which is better able to effectively distinguish pathogenic splicing-relevant variants from benign variants. S-CAP is a novel splicing pathogenicity predictor that reduces the number of splicing-relevant variants of uncertain significance in patient exomes by 41%, a nearly 3-fold improvement over existing noncoding pathogenicity measures while correctly classifying known pathogenic splicing-relevant variants with a clinical-grade 95% sensitivity.

genomics↗

AMELIE accelerates Mendelian patient diagnosis directly from the primary literature

The diagnosis of Mendelian disorders requires labor-intensive literature research. Our software system AMELIE (Automatic Mendelian Literature Evaluation) greatly automates this process. AMELIE parses hundreds of thousands of full text articles to find an underlying diagnosis to explain a patients phenotypes given the patients exome. AMELIE prioritizes patient candidate genes for their likelihood of causing the patients phenotypes. Diagnosis of singleton patients (without relatives exomes) is the most time-consuming scenario. AMELIEs gene ranking method was tested on 215 singleton Mendelian patients with a clinical diagnosis. AMELIE ranked the causal gene among the top 2 in the majority (63%) of cases. Examining AMELIEs top 10 genes, amounting to 8% of 124 candidate genes with rare functional variants per patient, results in diagnosis for 95% of cases. Strikingly, training only on gene pathogenicity knowledge from 2011 leads to identical performance compared to training on current data. An accompanying analysis web portal has launched at AMELIE.stanford.edu.

genetics↗

Revealing the causative variant in Mendelian patient genomes without revealing patient genomes

Given the rapidly growing utility of critical health information revealed in the human genome, secure genomic computation is essential to moving forward, especially as genome sequencing becomes commonplace. We devise and implement proof-of-principle computational operations for precisely identifying causal variants in Mendelian patients using secure multiparty computation methods based on Yaos protocol. We show multiple real scenarios (small patient cohorts, trio analysis, two hospital collaboration) where the causal variant is discovered jointly, while keeping up to 99.7% of all participants most sensitive genomic information private. All similar operations performed today to diagnose such cases are done openly, keeping 0% of participants genomic information private. Our work will help usher in an era where genomes can be both utilized and truly protected.

genomics↗