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Gade, S.

Publications and source records attributed to Gade, S..

3 recordsLinked to original sources

High-throughput platforms for genetic perturbation screening using CRISPR/Cas9 in human iPSC-derived macrophages for drug discovery

Human induced Pluripotent Stem Cell (hiPSC) models have revolutionised drug discovery, offering high translational relevance for recapitulating disease biology and thereby the potential to help reduce drug attrition. Macrophages are pivotal for maintaining tissue homeostasis and orchestrating immune responses; their dysregulation underpins several diseases, including autoinflammatory disorders, neurodegeneration, and cancer. Therapeutically targeting this cell type presents an attractive strategy to simultaneously influence multiple cellular mediators and functions. We have established a scalable, semi-automated, and physiologically relevant hiPSC-derived macrophage model, rigorously characterised through deep comparative multi-omics. We have also integrated our hiPSC-derived macrophage platform with large-scale CRISPR screening workflows designed for parallel genetic interrogation of thousands of gene targets, in both arrayed and pooled formats. In this manuscript, we apply those genetic screening methods to hiPSC-derived macrophages and showcase how genetic perturbations alter pro- and anti-inflammatory transcriptional signatures and significantly impact functional phenotypes in this cell model. This integrated approach allows for the exploration of novel genetic insights linked to causal disease biology, advancing myeloid cell-associated target discovery across a broad spectrum of therapeutic areas.

immunology↗

Discovery of an orally available potent ER aminopeptidase 1 (ERAP1) inhibitor that enhances anti-tumor responses and limits inflammatory autoimmunity in vivo.

Endoplasmic reticulum aminopeptidase 1 (ERAP1) is an intracellular enzyme that can regulate immune responses primarily by proteolytically processing peptides before loading and presentation on the cell surface by major histocompatibility class I molecules (MHC-I). ERAP1 activity can either reduce the immunogenicity of cancer cells by over-trimming cancer-associated antigenic peptides or contribute to autoimmunity by generating self-antigenic peptides. As a result, ERAP1 inhibition has emerged as a tractable approach for cancer immunotherapy and specific classes of autoimmunity. Here, we describe the discovery, after hit-to-lead optimization, of a potent and selective ERAP1 inhibitor based on the pyrrolidine 3-carboxylic acid scaffold that targets the regulatory allosteric site. The compound has favourable in vivo pharmacokinetics, including oral bioavailability, and can regulate the immunopeptidome of cancer cells and enhance cancer cell antigenicity in vivo in a dose-dependent manner, controlling tumor growth. In addition, when administered in the murine collagen-induced arthritis model, it does not induce any exacerbation of autoimmune responses but rather results in a dose-dependent therapeutic benefit. Our results demonstrate that ERAP1 inhibition can constitute a tractable approach to modulating immune responses for therapeutic applications, providing mechanistic insight and a valuable lead and in vivo tool for further drug development efforts and for interrogating ERAP1 biology.

immunology↗

Integration of Thermal Proteome Profiling with phosphoproteomic and transcriptomic data via mechanistic network models decodes the molecular response to PARP inhibition

The deregulation of complex diseases often spans multiple molecular processes. A multimodal functional characterization of these processes can shed light on the disease mechanisms and the effect of drugs. Thermal Proteome Profiling (TPP) is a mass-spectrometry based technique assessing changes in thermal protein stability that can serve as proxies of functional changes of the proteome. These unique insights of TPP can complement those obtained by other omics technologies. Here, we show how TPP can be integrated with phosphoproteomics and transcriptomics in a network-based approach using COSMOS, a framework for causal integration of multi-omics, to provide an integrated view of transcription factors, kinases and proteins with altered thermal stability. This allowed us to recover known mechanistic consequences of PARP inhibition in ovarian cancer cells on cell cycle and DNA damage response in detail and to uncover new insights into drug response mechanisms related to interferon and hippo signaling. We found that TPP complements the other omics data and allowed us to obtain a network model with higher coverage of the main underlying mechanisms. These results illustrate the added value of TPP, and more generally the power of network models to integrate the information provided by different omics technologies. We anticipate that this strategy can be used to broadly integrate functional proteomics with other omics to study complex molecular processes. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=63 SRC="FIGDIR/small/553354v1_ufig1.gif" ALT="Figure 1"> View larger version (18K): org.highwire.dtl.DTLVardef@af952eorg.highwire.dtl.DTLVardef@16b1d1borg.highwire.dtl.DTLVardef@1440787org.highwire.dtl.DTLVardef@14a4227_HPS_FORMAT_FIGEXP M_FIG C_FIG

systems biology↗