Human Liver Tissue Storage Methods For Multiomic Applications v1

Nakib, Diana

doi:10.17504/protocols.io.261ge34qdl47/v1

Cited by 2 publications

(1 citation statement)

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“…The copyright holder for this preprint this version posted August 1, 2023. ; https://doi.org/10.1101/2023.07. 28.550550 doi: bioRxiv preprint sections were snap frozen and stored in liquid nitrogen, or embedded in optimal cutting temperature (OCT) medium to be stored at -80C (23). Nuclei for single nucleus RNA sequencing (snRNAseq) (3 PSC explants) were extracted from snap frozen tissue as described previously (3,24).…”

Section: Ndd and Psc Sample Collection For Scrnaseq And Snrnaseqmentioning

confidence: 99%

Single-cell and spatial transcriptomics reveals the human liver immunological landscape and myeloid dysfunction in PSC

Andrews,

Nakib,

Perciani

et al. 2023

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Background: Primary sclerosing cholangitis (PSC) is a serious immune-mediated cholestatic liver disease characterized by bile retention, biliary tree destruction, and progressive fibrosis leading to end stage liver disease and transplantation. There is an unmet need to understand the cellular composition of the PSC liver and how it underlies disease pathogenesis. As such, we generated a comprehensive atlas of the PSC liver and a reference healthy liver dataset using multiple multi-omic modalities with functional validation. Methods: In this work, we employed single-cell (12,000 cells), single-nuclei (23,000 nuclei) and spatial transcriptomics (1 sample by 10x Visium and 3 samples with multi-region profiling by Nanostring GeoMx DSP) to profile the cellular ecosystem in 5 patients with PSC. Transcriptomic profiles were compared to 100k single cell transcriptomes and spatial transcriptomics controls from 24 healthy neurologically deceased donor (NDD) livers. Flow cytometry and intracellular cytokine staining was performed to validate PSC-specific differences in immune phenotype and function. Results: PSC explants with cirrhosis of the liver parenchyma and prominent periductal fibrosis were associated with a unique population of hepatocytes which transformed to a cholangiocyte-like phenotype. Those hepatocytes were surrounded by diverse immune cell populations, including monocyte-like macrophages, liver-resident and circulating natural killer (NK) cells. Cytokines released by inflamed cholangiocytes and fibrosis-resident hepatic stellate cells and endothelial cells recruited CD4+T-cells, dendritic cells, and neutrophils to PSC liver tissues. Tissue-resident macrophages, by contrast, were reduced in number and exhibited a dysfunctional inflammatory response to LPS and IFN-Ɣ stimulation. Conclusions: We present the first comprehensive atlas of the PSC liver and demonstrate hyper-activation and exhaustion-like phenotypes of myeloid cells and markers of chronic cytokine expression in late-stage PSC lesions.

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