Experimental Competitive Bone Marrow Transplant Assays

Hétu-Arbour, Roxann; Bouali, Sarah; Heinonen, Krista M.

doi:10.1007/978-1-0716-0810-4_12

Cited by 3 publications

(3 citation statements)

References 28 publications

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“…Considering differences seen in homeostatic conditions in Vangl2 Δ/Δ mice, we next evaluated HSC functionality under hematopoietic pressure. The gold-standard assay for HSC function is to evaluate their ability to reconstitute a lethally irradiated host so we conducted competitive bone marrow transplant assays as described ( Kwarteng and Heinonen, 2016 ; Hétu-Arbour et al, 2021a ) and followed post-transplant recovery by analyzing peripheral blood reconstitution for 16 weeks and BM recovery at 20–22 weeks by flow cytometry ( Figure 4A ). To evaluate the possible age-related role of Vangl2, we performed BM transplants with young (2-month-old) and adult (6-month-old) donors.…”

Section: Resultsmentioning

confidence: 99%

“…Donor (CD45.2 + ; C57BL/6 background, control and Vangl2 Δ/Δ ) and competitor (CD45.1 + ; B6.SJL) BM cells were analyzed by flow cytometry prior to transplant, and the quantities of total BM cells were adjusted to inject equivalent numbers of donor and competitor HSCs, similar to what has been previously described and as detailed below ( Kwarteng and Heinonen, 2016 ; Hétu-Arbour et al, 2021a ). Cells were pooled from two to three donors per experiment, and quantities of whole BM cells were normalized to the equivalent of 150 LT-HSCs (defined as CD150 + CD48 − Lin − Sca1 + c-Kit hi ).…”

Section: Methodsmentioning

confidence: 99%

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Vangl2 Promotes Hematopoietic Stem Cell Expansion

Bouali

Hétu-Arbour

Gardet

et al. 2022

Front. Cell Dev. Biol.

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Regulation of hematopoietic stem cell (HSC) self-renewal and differentiation is essential for their maintenance, and HSC polarity has been shown to play an important role in this regulation. Vangl2, a key component of the Wnt/polarity pathway, is expressed by fetal and adult HSCs, but its role in hematopoiesis and HSC function is unknown. Here we show the deletion of Vangl2 in mouse hematopoietic cells impairs HSC expansion and hematopoietic recovery post-transplant. Old Vangl2-deficient mice showed increased expansion of myeloid-biased multipotent progenitor cells concomitant with splenomegaly. Moreover, Vangl2-deficient cells were not able to effectively reconstitute the recipient bone marrow in serial transplants, or when coming from slightly older donors, demonstrating impaired self-renewal or expansion. Aged Vangl2-deficient HSCs displayed increased levels of cell cycle inhibitor p16INK4a and active β–catenin, which could contribute to their impaired function. Overall, our findings identify Vangl2 as a new regulator of hematopoiesis.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Vangl2 Promotes Hematopoietic Stem Cell Expansion

Bouali

Hétu-Arbour

Gardet

et al. 2022

Front. Cell Dev. Biol.

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“…Sex‐matched littermates were used as controls. For mixed BM chimeras, Vav‐iCre + Wnt4 fl/fl ( Wnt4 Δ/Δ ) or control BM cells (CD45.2 + ) were combined with 5 × 10 5 competitor cells (CD45.1 + or CD45.1 + /CD45.2 + ) in a 1:1 Lin − Sca‐1 + c‐Kit + (LSK) CD150 + cell equivalent ratio (as previously detailed 20,21 ) and injected into the lateral tail vein of lethally irradiated (2 × 450 rad) congenic recipient mice (CD45.1 + /CD45.2 + ). For lipopolysaccharide (LPS) stimulation assays, 1 dose of 1 mg/kg was administered by intraperitoneal injection into Vav‐iCre + Wnt4 fl/fl , LysMCre + Wnt4 fl/fl or control mice.…”

Section: Methodsmentioning

confidence: 99%

Cell-Intrinsic WNT4 Promotes Hematopoietic Stem and Progenitor Cell Self-Renewal

et al. 2021

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Although intracellular Wnt signaling pathways need to be tightly regulated to promote hematopoietic stem cell self-renewal, the source and identity of important Wnt ligands in the bone marrow is still largely unknown. The noncanonical ligand Wnt4 is expressed in the bone marrow as well as in the stroma, and its overexpression in fetal liver cells facilitates thymic recovery; however, its impact on adult hematopoietic stem cell function remains unclear. Here, we report that the deletion of Wnt4 from hematopoietic cells in mice (Wnt4Δ/Δ) resulted in decreased lymphopoiesis at steady state. This was likely at least in part due to the increased proinflammatory environment present in the bone marrow of Wnt4Δ/Δ mice. Wnt4Δ/Δ hematopoietic stem cells displayed reduced reconstitution capacity in serial transplants, thus demonstrating defective self-renewal, and they expanded poorly in response to lipopolysaccharide stimulation. This appeared to be the result of the absence of Wnt4 in stem/progenitor cells, as myeloid-restricted Wnt4 deletion had no notable effect. Finally, we observed that Wnt4Δ/Δ stem/progenitor cells were more quiescent, presenting enhanced levels of stress-associated JNK phosphorylation and p16INK4a expression, likely contributing to the reduced expansion observed in transplants. In conclusion, our results identify a new, largely autocrine role for Wnt4 in hematopoietic stem cell self-renewal, suggesting that regulation of Wnt signaling in hematopoiesis may not need Wnt secretion and could be independent of morphogen gradients.

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Systemic administration of a viral nanoparticle neoadjuvant prevents lung metastasis development through emergency myelopoiesis

Bourguignon,

Hétu-Arbour,

Charpentier

et al. 2024

OncoImmunology

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Cancer presents a significant public health concern, particularly in the context of metastatic disease. Surgical removal of primary tumors, while essential, can inadvertently heighten the risk of metastasis. Thus, there is a critical need for innovative neoadjuvant therapies capable of curtailing metastatic progression before or immediately following tumor resection. Addressing this imperative, the papaya mosaic virus nanoparticle (PapMV) has demonstrated potent immunostimulatory capabilities against both viruses and tumors, effectively hindering their proliferation. Our study reveals that PapMV exerts a protective effect against lung metastasis when administered systemically prior to tumor implantation or during the early stages of metastasis in various mouse models of cancer. This anti-tumor effect is initiated by the recruitment of myeloid cells in the lungs. These cells adopt a pro-inflammatory profile, secreting cytokines such as IFN-α, thus fostering a tumor microenvironment inhospitable to tumor progression. Crucially, this protective mechanism hinges on the presence of macrophages before treatment. TLR7 and IFN-I signaling pathways also play pivotal roles in this process. Furthermore, our findings demonstrate that PapMV triggers the activation of the bone marrow emergency response, which accounts for the influx of myeloid cells into the lungs. This study unveils a novel aspect of PapMV’s functionality. By bolstering the immune system, PapMV confers robust protection against metastasis at an early stage of disease progression. This discovery holds promise for therapeutic intervention, particularly as a preemptive measure prior to or just after surgical intervention.

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Experimental Competitive Bone Marrow Transplant Assays

Cited by 3 publications

References 28 publications

Vangl2 Promotes Hematopoietic Stem Cell Expansion

Vangl2 Promotes Hematopoietic Stem Cell Expansion

Cell-Intrinsic WNT4 Promotes Hematopoietic Stem and Progenitor Cell Self-Renewal

Systemic administration of a viral nanoparticle neoadjuvant prevents lung metastasis development through emergency myelopoiesis

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