NLRP1 in Bone Marrow Microenvironment Controls Hematopoietic Reconstitution After Transplantation

Abstract

Pretreatment before transplantation initiates an inflammatory response. Inflammasomes are key regulators of immune and inflammatory responses, but their role in regulating hematopoiesis is unclear. Our study intended to assess the role and mechanism of nucleotide-binding domain and leucine-rich repeat pyrin-domain containing protein 1 (NLRP1) in the bone marrow microenvironment on hematopoiesis regulation. To explore the effects of an absence of NLRP1 on hematopoietic reconstitution, we established a hematopoietic cell transplantation model by infusing bone marrow mononuclear cells of wild-type C57BL/6 mice into either NLRP1 knockout (NLRP1-KO) or wild-type C57BL/6 mice. Using the transplantation model, the role of NLRP1 in the bone marrow microenvironment was determined by flow cytometry, hemacytometry, and hematoxylin and eosin staining. As the major component of the bone marrow microenvironment, mesenchymal stem cells (MSCs) were isolated to analyze the effects of NLRP1 on them by osteogenic and adipogenic induction. Endothelial cells (ECs) were isolated and sorted by magnetic beads. The expression of adhesion molecules and their relationship with nuclear factor kappa B (NF-κB) were measured by immunofluorescence, enzyme-linked immunosorbent assay, and western blot. Finally, the effect of NLRP1-deleted MSCs or ECs on hematopoietic stem and progenitor cells (HSPCs) was examined by establishing co-culture models. Compared with C57BL/6 recipients, reduced inflammatory cell infiltration, decreased levels of proinflammatory cytokines interleukin (IL)-18, IL-1β, IL-6, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ), together with reduced pathological injury of bone marrow, were observed in NLRP1-KO recipients after transplantation. However, increased HSPC engraftment and hematopoietic reconstitution were detected in NLRP1-KO recipients after transplantation. Furthermore, MSCs isolated from NLRP1-KO mice had decreased osteogenic and adipogenic differentiation and increased proliferation and differentiation of HSPCs. The expression of adhesion molecules in ECs from NLRP1-KO mice was increased due to the promotion of nuclear translocation of NF-κB; these adhesion molecules are critical for hematopoietic stem cell homing. Knockout of NLRP1 in the bone marrow microenvironment could significantly relieve bone marrow inflammatory response and promote hematopoietic reconstitution, perhaps by regulating MSCs and ECs, indicating that NLRP1 might be a target for the treatment of delayed hematopoietic and immune recovery in patients after hematopoietic stem cell transplantation.