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Biology subjects

Malara, A.

Publications and source records attributed to Malara, A..

3 recordsLinked to original sources

Fibronectin silencing remodels the fibrotic bone marrow niche

Bone marrow (BM) fibrosis is a defining feature of Primary Myelofibrosis, yet the extracellular matrix (ECM) that sustains the fibrotic niche remains largely untargeted therapeutically. Fibronectin (FN), a key organizer of ECM architecture and growth factor signaling, accumulates in the fibrotic BM niche and correlates with disease severity. Here, we investigated whether systemic FN depletion can disrupt pathological remodeling of the BM niche. We developed a chemically modified antisense oligonucleotide targeting FN1 (FN ASO), which reduced FN expression and assembly and impaired transforming growth factor-{beta}1 (TGF-{beta}1)-induced activation of mesenchymal stromal cells. Systemic FN ASO administration efficiently depleted circulating and BM FN in mice and was well tolerated during short-term treatment. In two complementary models of BM fibrosis, FN silencing reduced FN matrix organization, stromal activation, active TGF{beta}-1, reticulin, and collagen deposition. In Jak2V617F-driven disease, transcriptomic profiling further revealed attenuation of ECM, TGF-{beta}1, and mesenchymal remodeling programs, accompanied by changes in inflammatory signaling. Collectively, these findings identify FN as an active organizer of the fibrotic BM niche and show that its systemic silencing uncouple ECM organization from profibrotic stromal signaling, providing rationale for targeting FN in myelofibrosis with broader therapeutic relevance to fibrosis-driven diseases.

cancer biology↗

A genetic anemia disentangles the dual effect of chronically reduced thrombopoietin on hematopoietic stem cells and bone marrow niche

Many studies unraveled the reciprocal regulation between hematopoietic stem cells (HSC) and the bone marrow (BM) niche by using transgenic knock-out or reporter mice, often overlooking the pathophysiological relevance and the existence of compensatory mechanisms. Here, we exploit the th3 murine model of a genetic anemia, beta-thalassemia (BThal), to investigate the dual impact of chronically reduced thrombopoietin (TPO) production on HSC and megakaryocytic BM niche. Decreased systemic TPO downregulates stemness programs in HSC and impairs their commitment towards megakaryocytes (Mk). We uncover dysmegakaryopoiesis in the BM from BThal patients and mice with defective Mk maturation and reduced production of niche factors, altering the HSC-niche crosstalk. Notably, in vivo stimulation of the TPO signaling restores HSC quiescence, Mk maturation and niche support. These findings demonstrate that TPO regulates HSC through both direct and niche-mediated routes, underscoring the value of using a genetic disease model to disentangle general molecular mechanisms.

cell biology↗

A bioprinted silk marrow niche reveals mechanical regulation of human megakaryopoiesis under genotoxic stress

Hematopoietic stem and progenitor cells (HSPCs) reside in a mechanically distinct bone marrow niche, yet how niche biomechanics shape genome stability and stress responses has been difficult to test because conventional two-dimensional (2D) culture lacks marrow viscoelasticity and uses surfaces that activate platelets, confounding hematopoietic readouts. Here, we show that this methodological gap has masked a basic principle: the marrow niche actively constrains genotoxic stress signaling in HSPCs, and 2D culture systematically overstates DNA damage and impairs differentiation in vitro. We engineered silk fibroin, a biologically inert biomaterial that does not activate platelets and recapitulates marrow viscoelasticity, into SilkInk, a 3D-bioprintable bioink, and used it to reconstruct a biomimetic marrow microenvironment. HSPCs encapsulated in SilkInk preserved clonogenic potential and multilineage differentiation, whereas 2D-cultured HSPCs activated cytoskeletal-tension and genome-surveillance programs characteristic of chronic stress, including pathways related to replication stress, DNA damage response, and redox stress. Cell phenotyping and single-cell RNA sequencing during megakaryopoiesis revealed that SilkInk supported coordinated endomitotic progression and terminal maturation, with progression from CD34+CD61-CD41-CD42b- progenitors to CD34-CD61+CD41+CD42b+ megakaryocytes, including increased 8N and >16N populations, whereas 2D culture and conventional 3D hydrogels sustained DNA damage signaling and impaired thrombopoiesis. The same hierarchy held under cytotoxic challenge, as 5-fluorouracil amplified DNA damage and crippled platelet output in 2D, whereas SilkInk-encapsulated HSPCs maintained differentiation, mirroring native marrow resilience. These findings reposition niche mechanics as an active determinant of hematopoietic genome stability and establish SilkInk as a physiologically faithful platform for studying hematopoiesis and predicting marrow responses to chemotherapy.

Cell Biology↗