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Biology subjects

Giglio, R. M.

Publications and source records attributed to Giglio, R. M..

5 recordsLinked to original sources

SHERLOCK: Structured representation learning and causal inference of downstream perturbation effects

Understanding the effects of genetic, molecular, and experimental perturbations is essential for decoding cellular mechanisms and guiding biomedical interventions. Existing computational approaches are typically designed for individual tasks, such as predicting perturbation responses, characterizing downstream transcriptional effects, or modeling perturbation combinations, and therefore do not provide a unified framework for learning interpretable perturbation representations while enabling causal analysis of downstream effects and characterization of condition-dependent responses. We present SHERLOCK, an interpretable deep generative framework for single-cell perturbation analysis that represents perturbation effects as structured interventions on a latent baseline cellular state. SHERLOCK learns correlated and sparse perturbation representations that organize genetic and pharmacological perturbations according to shared transcriptional responses. % By formulating perturbations as interventions within a structural causal model, it enables counterfactual estimation of their downstream transcriptional effects under explicit identifiability assumptions. The same framework quantifies how perturbation responses vary across conditions and compositionally models combinatorial perturbations, enabling prediction of held-out combinations and classification of genetic interactions. Across genome-scale CRISPR, chemical, and spatial perturbation datasets, SHERLOCK recovers perturbation relationships concordant with known biological pathways and pharmacological properties, identifies condition-dependent responses, and predicts combinatorial perturbation effects. Together, SHERLOCK provides a unified framework for interpretable and causal analysis of perturbation effects across diverse single-cell perturbation experiments.

genomics↗

Mapping kinase-dependent tumor immune adaptation with multiplexed single-cell CRISPR screens

Immune dysfunction in cancer is enacted by multiple programs, including tumor cell-intrinsic responses to distinct immune subpopulations. A subset of these immune evasion programs can be systematically recapitulated through direct tumor-immune interactions in vitro. Here, we present an integrated, high-throughput single-cell CRISPR screening framework focused on the protein kinome for mapping the tumor-intrinsic regulation of T cell-driven immune pressure in glioblastoma (GBM). We combine pooled CRISPR interference and activation (CRISPRi/a) with immune-matched NY-ESO-1 antigen-specific allogeneic GBM-T cell co-culture and massively multiplexed single-cell transcriptomics to systematically quantify how genetic perturbation reshapes baseline tumor state and adaptive responses across graded effector-to-target ratios. We further leverage deep generative models for analyzing pooled CRISPR screens to decipher the effects of genetic perturbations on the mechanisms of tumor resistance. This framework resolves distinct modules of immune evasion and survival, including the regulation of the antigen-presentation machinery, interferon/NF-{kappa}B signaling, oxidative stress resilience, and checkpoint/cytokine programs, while identifying perturbations that reroute the continuous tumor transcriptional trajectory induced by T cell engagement. A secondary chemical screen in patient-derived GBM cultures identified putative kinase targets of immune evasion phenotypes (e.g., EPHA2 and PDGFRA), whose inhibition leads to the blockade of evasive programs and enhances T cell-mediated GBM killing. Together, this workflow provides a scalable blueprint for comprehensive charting of the genetic control of tumor-immune interactions.

genomics↗

Multiplexed single-cell transcriptomics reveals diverse phenotypic outcomes for pathogenic SHP2 variants

The protein tyrosine phosphatase SHP2, encoded by PTPN11, is an important regulator of Ras/MAPK signaling that acts downstream of receptor tyrosine kinases and other transmembrane receptors. Germline PTPN11 mutations cause developmental disorders such as Noonan Syndrome, whereas somatic mutations drive various cancers. While many pathogenic mutations enhance SHP2 catalytic activity, others are inactivating or affect protein interactions, confounding our understanding of SHP2-driven disease. Here, we combine single-cell transcriptional profiling of cells expressing clinically diverse SHP2 variants with protein biochemistry, structural analysis, and cell biology to explain how pathogenic mutations dysregulate signaling. Our analyses reveal that loss of catalytic activity does not phenocopy SHP2 knock-out at the gene expression level, that some mechanistically distinct mutations have convergent phenotypic effects, and that different mutations at the same hotspot residue can yield divergent cell states. These findings provide a framework for understanding the connection between SHP2 structural perturbations, cellular outcomes, and human diseases.

molecular biology↗

Lineage-specific proteome remodeling of diverse lung cancer cells by targeted epigenetic inhibitors

Epigenetic inhibitors exhibit powerful antiproliferative and anticancer activities. However, cellular responses to small-molecule epigenetic inhibition are heterogeneous and dependent on factors such as the genetic background and metabolic state of cells, as well as on-/off-target engagement of individual small-molecule compounds. The molecular study of the extent of this heterogeneity often measures changes in a single cell line. To more comprehensively profile the effects of small-molecule perturbations and their influence on heterogeneous cellular responses, we present a molecular resource based on the quantification of chromatin, proteome, and transcriptome remodeling due to histone deacetylase inhibitors (HDACi) in non-isogenic cell lines. Through quantitative molecular profiling of 10,621 proteins, these data reveal coordinated molecular remodeling of HDACi treated cancer cells. HDACi-regulated proteins differ greatly across cell lines with consistent (JUN, MAP2K3, CDKN1A) and divergent (CCND3, ASF1B, BRD7) cell-state effectors. Together these data provide valuable insight into cell-type driven and heterogeneous responses that must be taken into consideration when monitoring molecular perturbations in culture models. We have also built a web interface for the extensive amount of data to allow users to explore the data as a resource for understanding chemical perturbation of diverse cell types.

cancer biology↗

A heterogenous pharmaco-transcriptomic landscape induced by targeting a single oncogenic kinase

Over-activation of the epidermal growth factor receptor (EGFR) is a hallmark of glioblastoma. However, EGFR-targeted therapies have led to minimal clinical response. While delivery of EGFR inhibitors (EGFRis) to the brain constitutes a major challenge, how additional drug-specific features alter efficacy remains poorly understood. We introduce SCHEMATIC, which integrates multiplex single-cell chemical transcriptomics with deep-generative classification to resolve chemotype-specific and shared programs and apply it to to define the molecular response of glioblastoma to EGFRis. We identify programs that differ by the chemical properties of EGFRis, including induction of adaptive transcription and modulation of immunogenic gene expression. We find that induction of an adaptive transcriptional program is associated with persistence of surviving cells after EGFR inhibition, and that concurrent EGFR/PI3K inhibition attenuates this program. We also find that pro-immunogenic expression changes associated with a subset of tyrphostin-family EGFR inhibitors are accompanied by enhanced antigen-specific cytotoxic T-cell killing in vitro. Our study provides a framework that considers each agents unique and often unknown poly-pharmacology to prioritize compounds pre-clinically that induce favorable molecular responses.

genomics↗