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Understanding Heterogeneity of Human Bone Marrow Plasma Cell Maturation and Survival Pathways by Single Cell Analyses

ABSTRACT: Understanding Heterogeneity of Human Bone Marrow Plasma Cell Maturation and Survival Pathways by Single Cell Analyses

PROVIDER: PRJNA962176 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24318900.fastq.gz	Fastqsanger.gz
	SRR24319000.fastq.gz	Fastqsanger.gz
	SRR24319100.fastq.gz	Fastqsanger.gz
	SRR24318901.fastq.gz	Fastqsanger.gz
	SRR24319001.fastq.gz	Fastqsanger.gz

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Understanding Heterogeneity of Human Bone Marrow Plasma Cell Maturation and Survival Pathways by Single Cell Analyses [scVDJ-seq]

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Understanding Heterogeneity of Human Bone Marrow Plasma Cell Maturation and Survival Pathways by Single Cell Analyses [scVDJ-seq]

Project description:Human bone marrow (BM) plasma cells are heterogeneous, ranging from newly arrived antibody-secreting cells (ASC) to long-lived plasma cells (LLPC). We provide single cell transcriptional resolution of 17,347 BM ASC from 5 healthy adults. Fifteen clusters were identified ranging from newly minted ASC (cluster 1) expressing MKI67 and high MHC Class II that progressed to late clusters 5-8 through intermediate clusters 2-4. Additional clusters included IgM-predominant ASC of likely extra-follicular origin; IFN-responsive; and high mitochondrial activity ASC. Late ASCs were distinguished by differences in G2M checkpoints, MTOR signaling, distinct metabolic pathways, CD38 expression, and utilization of TNF-receptor superfamily members. They mature through two distinct paths differentiated by the degree of TNF signaling through NFKB. This study provides the first single cell resolution atlas and molecular roadmap of LLPC maturation, thereby providing insight into differentiation trajectories and molecular regulation of these essential processes in the human BM microniche. This information enables investigation of the origin of protective and pathogenic antibodies in multiple diseases and development of new strategies targeted to the enhancement or depletion of the corresponding ASC.

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Project description:Plasma cells provide protective antibodies following an infection or vaccination. A network of intrinsic and extrinsic factors fine-tunes the generation of a heterogenous plasma cell pool with varying metabolic requirements, transcriptional profiles and lifespans. Among these intrinsic factors, the B cell maturation antigen (BCMA) has been implicated in APRIL-mediated survival of long-lived plasma cells. Therefore, we introduced a tdTomato reporter into the Tnfrsf17 (BCMA) locus, to decode the developmental steps behind the generation of mature plasma cell subsets. We demonstrate that the BCMA:Tom reporter is exclusively detected in plasma cells, with its expression differing between IgH isotypes and increasing with plasma cell maturity. Combined with a Blimp1-GFP reporter, the BCMA:Tom reporter allowed us to track the step-wise maturation from newly formed to late plasma cells in various plasma cell subpopulations. Notably, in vitro-stimulated B cells induce BCMA only after adoptive transfer into recipient mice, emphasizing the impact of the in vivo microenvironment in establishing a mature plasma cell compartment. The BCMA:Tom reporter mouse provides a robust tool for tracking plasma cell development and maturation with flow cytometry or advanced imaging techniques, enabling a deeper understanding of the mechanisms regulating plasma cell heterogeneity and longevity.

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Project description:Survival of antibody-secreting plasma cells is a prerequisite to establish long-lasting humoral immune protection. The B cell maturation antigen (BCMA) has been proposed to promote APRIL-mediated survival signals in plasma cells. However, the extensive shedding of murine BCMA from the plasma cell surface raises doubts about its ability to function as a signaling receptor. To unequivocally establish whether BCMA is required for long-lived plasma cell survival, we established two independent BCMA-deficient mouse lines and analyzed the survival of antigen-specific plasma cells after immunization. In contrast to published data, we found comparable numbers of long-lived antigen-specific plasma cells after primary and secondary protein and mRNA immunizations in BCMA-deficient mice. In addition, wildtype and BCMA-deficient plasma cells showed comparable transcriptome profiles with no indication for reduced survival signaling. Interestingly, total plasma cell numbers were increased after boost immunization in bone marrow and mesenteric lymph nodes of BCMA-deficient mice. Therefore, BCMA has no intrinsic role in maintaining long-lived plasma cells. We propose a function of BCMA limited to acting as a soluble decoy receptor for APRIL and fine-tuneing the size of the plasma cell population by limiting the availability of survival factors. This finding removes a piece of the puzzle of the mechanisms that contribute to long-lived plasma cell survival.

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