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Single-cell RNA sequencing of murine Colon26 tumors after combination treatment with HMGN1 immunostimulatory peptide (minP1) with anti-PD-L1 antibody

ABSTRACT: Single-cell RNA sequencing of murine Colon26 tumors after combination treatment with HMGN1 immunostimulatory peptide (minP1) with anti-PD-L1 antibody

PROVIDER: PRJNA703835 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13761518_1.fastq.gz	Fastqsanger.gz
	SRR13761518_2.fastq.gz	Fastqsanger.gz
	SRR13761519_1.fastq.gz	Fastqsanger.gz
	SRR13761519_2.fastq.gz	Fastqsanger.gz

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Systemically administered HMGN1 immunostimulatory peptide with anti-PD-L1 antibody exerts synergistic antitumor effects

Project description:In this study, we explore a new combination therapy of anti-PD-L1 antibody with an immunostimulatory peptide derived from an alarmin high mobility group nucleosome binding protein 1 (HMGN1). Combination treatement of HMGN1 immunostimulatory peptide with anti-PD-L1 antibody exerted robust anti-tumor effects in Colon26 subtaneous murine model. To understand transcriptomic differences of immune cell population in the tumor-bearing mice after treated with single-agent or combination therapy of HMGN1 with anti-PD-L1 antibody, we performed transcriptome analysis on tumor tissue using 5’end Serial Analysis of Gene Expression (SAGE)-sequencing with an Ion 540 Chef kit, an Ion 540 Chip kit, and an Ion S5 Sequencer (Thermo Fisher Scientific).

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Single-cell RNA sequencing of murine Colon26 tumors after combination treatment with HMGN1 immunostimulatory peptide (minP1) with anti-PD-L1 antibody

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Systemically administered HMGN1 immunostimulatory peptide with anti-PD-L1 antibody exerts synergistic antitumor effects

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Rapid Proteomic Screen of CRISPR Experiment Outcome by Data Independent Acquisition Mass Spectrometry: A Case Study for HMGN1

Project description:CRISPR-Cas gene editing holds substantial promise in many biomedical disciplines and basic research. Due to the important functional implications of non-histone chromosomal protein HMG-14 (HMGN1) in regulating chromatin structure and tumor immunity, we performed gene knockout of HMGN1 by CRISPR in cancer cells and studied the following proteomic regulation events. In particular we utilized DIA mass spectrometry (DIA-MS) and reproducibly measured more than 6200 proteins (protein- FDR 1%) and more than 82,000 peptide precursors in the single MS shots of two hours. HMGN1 protein deletion was confidently verified in all of the clone- and dish- replicates following CRISPR by DIA-MS. Statistical analysis revealed 144 proteins changed their expressions significantly after HMGN1 knockout. Functional annotation and enrichment analysis indicate the deletion of HMGN1 induces the histone inactivation, various stress pathways, remodeling of extracellular proteomes, and immune regulation processes related to complement and coagulation cascade and interferon alpha/ gamma response in cancer cells. These results shed new lights on the cellular functions of HMGN1. We suggest that DIA-MS can be reliably used as a rapid, robust, and cost-effective proteomic-screening tool to assess the outcome of the CRISPR experiments.

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