Periosteal skeletal stem cells can migrate into the bone marrow and support hematopoiesis after injury
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Curated by eLife
eLife Assessment
The study is valuable to the field, introducing a new model to test BM-periosteal stem cell function in vivo. The authors' findings suggested that periosteal stem cells are linked to hematopoietic regeneration. More comparisons with the conventional model and direct examination of periosteal stem cell factors in hematopoietic regeneration are missing. The observations are solid, however, the limitations in their experimental model made the overall impact incomplete; there is potential for improvements to be made in this area.
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Abstract
Skeletal stem cells have been isolated from various tissues, including periosteum and bone marrow, where they exhibit key functions in bone biology and hematopoiesis, respectively. The role of periosteal skeletal stem cells in bone regeneration and healing has been extensively studied, but their ability to contribute to the bone marrow stroma is still under debate. In the present study, we characterized a whole bone transplantation model that mimics the initial bone marrow necrosis and fatty infiltration seen after injury. Using this model and a lineage tracing approach, we observed the migration of periosteal skeletal stem cells into the bone marrow after transplantation. Once in the bone marrow, periosteal skeletal stem cells are phenotypically and functionally reprogrammed into bone marrow mesenchymal stem cells that express high levels of hematopoietic stem cell niche factors such as Cxcl12 and Kitl. In addition, using in-vitro and in-vivo approaches, we found that periosteal skeletal stem cells are more resistant to acute stress than bone marrow mesenchymal stem cells. These results highlight the plasticity of periosteal skeletal stem cells and their potential role in bone marrow regeneration after bone marrow injury.
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eLife Assessment
The study is valuable to the field, introducing a new model to test BM-periosteal stem cell function in vivo. The authors' findings suggested that periosteal stem cells are linked to hematopoietic regeneration. More comparisons with the conventional model and direct examination of periosteal stem cell factors in hematopoietic regeneration are missing. The observations are solid, however, the limitations in their experimental model made the overall impact incomplete; there is potential for improvements to be made in this area.
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Reviewer #1 (Public review):
The manuscript under review investigates the role of periosteal stem cells (P-SSC) in bone marrow regeneration using a whole-bone subcutaneous transplantation model. While the model is somewhat artificial, the findings were interesting, suggesting the migration of periosteal stem cells into the bone marrow and their potential to become bone marrow stromal cells. This indicates a significant plasticity of P-SSC consistent with previous reports using fracture models (Cell Stem Cell 29:1547, Dev Cell 59:1192).
Major Concerns
(1) The authors assert that the periosteal layer was completely removed in their model, which is crucial for their conclusions. To substantiate this claim, it is recommended that the authors provide evidence of the successful removal of the entire periosteal stem cell (P-SSC) population. A …
Reviewer #1 (Public review):
The manuscript under review investigates the role of periosteal stem cells (P-SSC) in bone marrow regeneration using a whole-bone subcutaneous transplantation model. While the model is somewhat artificial, the findings were interesting, suggesting the migration of periosteal stem cells into the bone marrow and their potential to become bone marrow stromal cells. This indicates a significant plasticity of P-SSC consistent with previous reports using fracture models (Cell Stem Cell 29:1547, Dev Cell 59:1192).
Major Concerns
(1) The authors assert that the periosteal layer was completely removed in their model, which is crucial for their conclusions. To substantiate this claim, it is recommended that the authors provide evidence of the successful removal of the entire periosteal stem cell (P-SSC) population. A colony-forming assay, with and without periosteal removal, could serve as a suitable method to demonstrate this.
(2) The observation that P-SSCs do not express Kitl or Cxcl12, while their bone marrow stromal cell (BM-MSC) derivatives do, is a key finding. To strengthen this conclusion, the authors are encouraged to repeat the experiment using Cxcl12 or Scf reporter alleles. Immunofluorescence staining that confirms the migration of periosteal cells and their transformation into Cxcl12- or Scf-reporter-positive cells would significantly enhance the paper's key conclusion.
(3) On page 8, line 20, the authors' statement regarding the detection of Periostin+ cells outside the periosteum layer could be misinterpreted due to the use of the periostin antibody. Given that periostin is an extracellular matrix protein, the staining may not accurately represent Periostin-expressing cells but rather the presence of periostin in the extracellular matrix. The authors should revise this section for greater precision.
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Reviewer #2 (Public review):
Summary:
The authors have established a femur graft model that allows the study of hematopoietic regeneration following transplantation. They have extensively characterized this model, demonstrating the loss of hematopoietic cells from the donor femur following transplantation, with recovery of hematopoiesis from recipient cells. They also show evidence that BM MSCs present in the graft following transplantation are graft-derived. They have utilized this model to show that following transplantation, periosteal cells respond by first expanding, then giving rise to more periosteal SSCs, and then migrating into the marrow to give rise to BM MSCs.
Strengths:
These studies are notable in several ways:
(1) Establishment of a novel femur graft model for the study of hematopoiesis;
(2) Use of lineage tracing and …
Reviewer #2 (Public review):
Summary:
The authors have established a femur graft model that allows the study of hematopoietic regeneration following transplantation. They have extensively characterized this model, demonstrating the loss of hematopoietic cells from the donor femur following transplantation, with recovery of hematopoiesis from recipient cells. They also show evidence that BM MSCs present in the graft following transplantation are graft-derived. They have utilized this model to show that following transplantation, periosteal cells respond by first expanding, then giving rise to more periosteal SSCs, and then migrating into the marrow to give rise to BM MSCs.
Strengths:
These studies are notable in several ways:
(1) Establishment of a novel femur graft model for the study of hematopoiesis;
(2) Use of lineage tracing and surgery models to demonstrate that periosteal cells can give rise to BM MSCs.
Weaknesses:
There are a few weaknesses. First, the authors do not definitively demonstrate the requirement of periosteal SSC movement into the BM cavity for hematopoietic recovery. Hematopoiesis recovers significantly before 5 months, even before significant P-SSC movement has been shown, and hematopoiesis recovers significantly even when periosteum has been stripped. Second, it is not clear how the periosteum is changing in the grafts. Which cells are expanding is unclear, and it is not clear if these cells have already adopted a more MSC-like phenotype prior to entering the marrow space. Indeed, given the presence of host-derived endothelial cells in the BM, these studies are reminiscent of prior studies from this group and others that re-endothelialization of the marrow may be much more important for determining hematopoietic regeneration, rather than the P-SSC migration. Third, the studies exploring the preferential depletion of BM MSCs vs P-SSCs are difficult to interpret. The single metabolic stress condition chosen was not well-justified, and the use of purified cell populations to study response to stress ex vivo may have introduced artifacts into the system.
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Reviewer #3 (Public review):
Summary:
Marchand, Akinnola, et al. describe the use of the novel model to study BM regeneration. Here, they harvest intact femurs and subcutaneously graft them into recipient mice. Similar to standard BM regeneration models, there is a rapid decrease in cellularity followed by a gradual recovery over 5 months within the grafts. At 5 months, these grafts have robust HSC activity, similar to HSCs isolated from the host femur. They find that periosteum skeletal stem cells (p-SSCs) are the primary source of BM-MSCs within the grafted femur and that these cells are more resistant to the acute stress of grafting the femur.
Strengths:
This is an interesting manuscript that describes a novel model to study BM regeneration. The model has tremendous promise.
Weaknesses:
The authors claim that grafting intact femurs …
Reviewer #3 (Public review):
Summary:
Marchand, Akinnola, et al. describe the use of the novel model to study BM regeneration. Here, they harvest intact femurs and subcutaneously graft them into recipient mice. Similar to standard BM regeneration models, there is a rapid decrease in cellularity followed by a gradual recovery over 5 months within the grafts. At 5 months, these grafts have robust HSC activity, similar to HSCs isolated from the host femur. They find that periosteum skeletal stem cells (p-SSCs) are the primary source of BM-MSCs within the grafted femur and that these cells are more resistant to the acute stress of grafting the femur.
Strengths:
This is an interesting manuscript that describes a novel model to study BM regeneration. The model has tremendous promise.
Weaknesses:
The authors claim that grafting intact femurs subcutaneously is a model of BM regeneration and can be used as a replacement for gold standard BM regeneration assays such as sublethal chemo/irradiation. However, there isn't enough explanation as to how this model is equivalent or superior to the traditional models. For instance, the authors claim that this model allows for the study of "BM regeneration in vivo in response to acute injury using genetic tools." This can and has been done numerous times with established, physiologically relevant BM regeneration models. The onus is on the authors to discuss or perform the necessary experiments to justify the use of this model. For example, standard BM regeneration models involve systemic damage that is akin to therapies that require BM regeneration. How is studying the current model that provides only an acute injury more relevant and useful than other models? As it stands, it seems as if the authors could have done all the experiments demonstrating the importance of these p-SSCs in the traditional myelosuppressive BM regeneration models to be more physiologically relevant. Along these lines, the use of a standard BM regeneration model (e.g., sublethal chemo/irradiation) as a critical control is missing and should be included. Even if the control doesn't demonstrate that p-SSCs can contribute to the BM-MSC during regeneration, it will still be important because it could be the justification for using the described model to specifically study p-SSCs' regulation of BM regeneration.
The authors perform some analysis that suggests that grafting a whole femur mimics BM regeneration, but there are many experiments missing from the manuscript that will be necessary to support the use of this model. To demonstrate that this new model mimics current BM regeneration models, the authors need to perform a careful examination of the early kinetics of hematopoietic recovery post-transplant. Complete blood counts should be performed on the grafts, focusing on white blood cells (particularly neutrophils), red blood cells, platelets, all critical indicators of BM regeneration. This analysis should be done at early time points that include weekly analysis for a minimum of 28 days following the graft. Additionally, understanding how and when the vasculature recovers is critical. This is particularly important because it is well-established that if there is a delay in vascular recovery, there is a delay in hematopoietic recovery. As mentioned above, a standard BM regeneration model should be used as a control.
The contribution of donor and host cells to the BM regeneration of the graft is interesting. Particularly, the chimerism of the vasculature. One can assume that for the graft to undergo BM regeneration, there needs to be the delivery of nutrients into the graft via the vasculature. The chimerism of the vascular network suggests that host endothelial cells anastomose with the graft. Host mice should have their vascular system labeled with a dye such as dextran to determine if anastomosis has occurred. If not, the authors need to explain how this graft survives up to 5 months. If anastomosis does occur, then it is very surprising that the hematopoietic system of the graft is not a chimera because this would essentially be a parabiosis model. This needs to be explained.
Most of the data presented for the resistance of p-SSCs to stress suggests DNA damage response. Do p-SSCs demonstrate a higher ability to resolve DNA damage? Do they accumulate less DNA damage? Staining for DNA damage foci or performing comet assays could be done to further define the mechanism of stress resistance properties of p-SSCs.
Given the importance of BM-MSCs in hematopoiesis and that the majority of the emerging BM-MSCs appear to be derived from p-SSCs, the authors should perform experiments to determine if p-SSC-derived BM-MSCs are critical regulators of BM regeneration. For example, the authors could test this by crossing the Postn-creER mice with iDTR mice to ablate these cells and see if recovery is inhibited or delayed. This should be done with the described periosteum-wrapped femur graft model as well as a control BM regeneration model. Demonstrating that the deletion of these cells affects BM regeneration in both models would further justify the physiological relevance and utility of the femur graft model.
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