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AMULET: a novel read count-based method for effective multiplet detection from single nucleus ATAC-seq data.

ABSTRACT: Detecting multiplets in single nucleus (sn)ATAC-seq data is challenging due to data sparsity and limited dynamic range. AMULET (ATAC-seq MULtiplet Estimation Tool) enumerates regions with greater than two uniquely aligned reads across the genome to effectively detect multiplets. We evaluate the method by generating snATAC-seq data in the human blood and pancreatic islet samples. AMULET has high precision, estimated via donor-based multiplexing, and high recall, estimated via simulated multiplets, compared to alternatives and identifies multiplets most effectively when a certain read depth of 25K median valid reads per nucleus is achieved.

SUBMITTER: Thibodeau A

PROVIDER: S-EPMC8408950 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Similar to other droplet-based single cell assays, single nucleus ATAC-seq (snATAC-seq) data harbor multiplets that confound downstream analyses. Detecting multiplets in snATAC-seq data is particularly challenging due to data sparsity and limited dynamic range (0 reads: closed chromatin, 1: open on in one parental chromosome allele, 2: open on in both alleles chromosomes). Yet, these unique data features offer an opportunity to identify multiplets. ATAC-DoubletDetector (https://ucarlab.github.io/ATAC-DoubletDetector/) AMULET (Atac MULtiplet Estimation Tool) exploits these unique features to detect multiplets by studying enumerates the number of regions with >2 uniquely aligned reads across the genome to effectively detect multiplets - an effective alternative to methods based on artificially-generated multiplets. We evaluated the method by generating snATAC-seq data (e.g., state-of-the-art ArchR). For benchmarking we generated snATAC-seq data and generated data fromeasured the efficacy of AMULET inm in two primary human tissues: peripheral human blood mononuclear cells (PBMCs) and pancreatic islet samples. AMULET detects had high multiplets with an estimated precision (estimated via donor-based multiplexing) and high recall (estimated via simulated doublets) compared to alternatives 0.57 precision and achieves 0.85 recall. When and was the most effective when a certain read depth is achieved (a certain read depth per nucleus is achieved samples are sequenced deeply (e.g., median read count per nucleus >20K25K) reads per nucleus in PBMCs), ATAC-DoubletDetector captured 85% of simulated doublets (i.e., recall), significantly outperforming ArchR (24%). For lower read depth, ATAC-DoubletDetector and ArchR produced complementary results. Moreover, ATAC-DoubletDetector was equally effective in identifying homotypic multiplets (i.e., multiplets from the same cell type), which are missed by simulation-based methods. Cell-specific marker peaks enabled accurate (85%) tracing of cellular origins of snATAC-seq multiplets. Accordingly, more abundant cells within a tissue are more likely to form multiplets and the majority of multiplets are homotypic. ATAC-DoubletDetector is a fast and effective multiplet detection/annotation tool for improved single cell epigenomic data analyses across diverse biological systems and conditions.

Dataset Information

AMULET: a novel read count-based method for effective multiplet detection from single nucleus ATAC-seq data.

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