GERDA datasets including NGS and SGA data
Description
Datasets linked to publication "Revealing viral and cellular dynamics of HIV-1 at the single-cell level during early treatment periods", Otte et al 2023 published in Cell Reports Methods pre-ART (antiretroviral therapy) cryo-conserved and and whole blood specimen were sampled for HIV-1 virus reservoir determination in HIV-1 positive individuals from the Swiss HIV Study Cohort. Patients were monitored for proviral (DNA), poly-A transcripts (RNA), late protein translation (Gag and Envelope reactivation co-detection assay, GERDA) and intact viruses (golden standard: viral outgrowth assay, VOA). In this dataset we deposited the pipeline for the multidimensional data analysis of our newly established GERDA method, using DBScan and tSNE. For further comprehension NGS and Sanger sequencing data were attached as processed and raw data (GenBank). Resubmitted to Cell Reports Methods (Jan-2023), accepted in principal (Mar-2023) GERDA is a new detection method to decipher the HIV-1 cellular reservoir in blood (tissue or any other specimen). It integrates HIV-1 Gag and Env co-detection along with cellular surface markers to reveal 1) what cells still contain HIV-1 translation competent virus and 2) which marker the respective infected cells express. The phenotypic marker repertoire of the cells allow to make predictions on potential homing and to assess the HIV-1 (tissue) reservoir. All FACS data were acquired on a LSRFortessa BD FACS machine (markers: CCR7, CD45RA, CD28, CD4, CD25, PD1, IntegrinB7, CLA, HIV-1 Env, HIV-1 Gag) Raw FACS data (pre-gated CD4CD3+ T-cells) were arcsin transformed and dimensionally reduced using optsne. Data was further clustered using DBSCAN and either individual clusters were further analyzed for individual marker expression or expression profiles of all relevant clusters were analyzed by heatmaps. Sequences before/after therapy initiation and during viral outgrowth cultures were monitored for individuals P01-46 and P04-56 by Next-generation sequencing (NGS of HIV-1 Envelope V3 loop only) and by Sanger (single genome amplification, SGA) data normalization code (by Julian Spagnuolo) FACS normalized data as CSV (XXX_arcsin.csv) OMIQ conText file (_OMIQ-context_XXX) arcsin normalized FACS data after optsne dimension reduction with OMIQ.ai as CSV file (XXXarcsin.csv.csv) R pipeline with codes (XXX_commented.R) P01_46-NGS and Sanger sequences P04_56-NGS and Sanger sequences
Files
Steps to reproduce
For all three datasets a step-by-step R coding pipeline is attached as well as R working environments for comparison.