Architecture of androgen receptor pathways amplifying glucagon-like peptide-1 insulinotropic action in male pancreatic β-cells

Table of Contents

What is this?

This repository contains coding scripts utilized for the analysis performed in the "Architecture of androgen receptor pathways amplifying glucagon-like peptide-1 insulinotropic action in male pancreatic β-cellse" publication (Xu-Qadir et. al, 2023). The purpose of providing the code here is to allow for transparency and robust data-analysis reproducibility. Most of the steps used for data analysis and visualization have been optimised for an average computing environment (for the year 2023). Some analyses however, require a high-performace computing environment (see computing environment). The methodology has already been described extensively in the manuscript. However, this analysis relies heavily on powerful scRNAseq analysis algorithms developed by the Satija lab, namely Seurat (Butler et al., 2018: Nature Biotechnology; Stuart et al., 2018: Cell) (for a complete list of dependencies and code utilized see analysis & visualization programs).

How can I use this data, and where can I find it?

Downloading Data files

Data files utilized in this analysis have been deposited in the Gene Expression Omnibus (GEO), gene expression data repository at the NIH. Data are part of the GSE131886 high-thoroughput sequencing repository and can be found here. Contact lead author for seurat object.

Data sub-structure

We povide raw FASTQ files generated from single-cell cDNA libraries sequenced by the Illumina sequencing platform, along with unfiltered post-alignment count files generated by the Cellranger software. In addition we also provide a gene expression matrix containing data on filtered gene counts across our dataset.

FASTQ files

These are sequencing reads generated by the Illumina sequencing platform. Files contain raw reads and sequencing efficiency information. These are the input files for the Cellranger software. and can be found here: GSE201256

Cellranger output (processed gene-counts of single cells/barcodes)

This contains data outputs of Cellranger, which was run using default settings. Code used to analyze data is a part of this repository. This data contains filtered/unfiltered count files for gene expression across barcodes/cells.

Data analysis of this dataset

Preliminary data-analyses involving n=3 ctrl + n=3 DHT Tx deidentified human cadaveric Islet samples.

Analysis and visualization programs

Cellranger software from 10X Genomics:

  1. Cellranger

R and R's integrated developmental environment RStudio:

  1. R v4.2.2 (x64 bit)
  2. RStudio build #386 (x64 bit)
  3. RTools v4.2.X
  4. Tutorial for R
  5. Tutorial for RStudio

scRNAseq analysis pipeline SEURAT developed by the Satija lab:

  1. Source code for Seurat
  2. Tutorials for Seurat

Setting up the right environment

  1. Install R
  2. Install Rstudio
  3. Once you have installed R and RStudio, you can run the script.
  4. If you need help understanding how commands are run in R use the [ctrl + enter] command or please visit here.
  5. If you run into problems, please open a new issue, you can do this by going to 'issues' and clicking on the 'new issue' icon.

Computing environment

Hardware and OS environment for running analysis

Environment

  1. Processor: Intel core i9-9900 (16cores x 16 threads)
  2. RAM: 128GB DDR3
  3. OS: Windows 10 Enterprise (x64 bit)

Citation

Contributors

  1. Mirza Muhammad Fahd Qadir - Github - to contact please Email

Lead Contacts

  1. Dr. Franck Mauvais-Jarvis MD PhD. - Dept of Medicine, Tulane University - to contact please Email

Acknowledgements

  1. NIDDK
  2. VA

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