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Arthritis augments breast cancer metastasis: role of mast cells and SCF/c-Kit signaling.

Das Roy L, Curry JM, Sahraei M, Besmer DM, Kidiyoor A, Gruber HE, Mukherjee P - Breast Cancer Res. (2013)

Bottom Line: We recently reported that mice with autoimmune arthritis have significantly increased incidence of bone and lung metastasis and decreased survival associated with breast cancer.This may primarily be due to the interaction of c-Kit receptor present on mast cells and stem cell factor (SCF, the ligand for ckit) expressed on tumor cells.Finally, we showed that targeting the SCF/cKit interaction with an anti-ckit antibody reduces the differentiation of mast cells and consequently reduces metastasis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Breast cancer remains the second leading cause of cancer-related deaths for women in the United States. Metastasis is regulated not only by intrinsic genetic changes in malignant cells, but also by the microenvironment, especially those associated with chronic inflammation. We recently reported that mice with autoimmune arthritis have significantly increased incidence of bone and lung metastasis and decreased survival associated with breast cancer. In this study, we evaluated the mechanism underlying the increased metastasis.

Methods: We used two mouse models; one that develops spontaneous autoimmune arthritis (SKG mice) injected with metastatic breast cancer cells (4T1), and another that develops spontaneous breast cancer (MMTV-PyV MT mice) injected with type II collagen to induce autoimmune arthritis. Mast cell levels and metastasis were monitored.

Results: First, we confirmed that breast tumor-bearing arthritic mice have a significantly higher incidence of bone and lung metastasis than do their nonarthritic counterparts. Next, we showed increased recruitment of mast cells within the primary tumor of arthritic mice, which facilitates metastasis. Next, we report that arthritic mice without any tumors have higher numbers of mast cells in the bones and lungs, which may be the underlying cause for the enhanced lung and bone metastases observed in the arthritic mice. Next, we showed that once the tumor cells populate the metastatic niches (bones and lungs), they further increase the mast cell population within the niche and assist in enhancing metastasis. This may primarily be due to the interaction of c-Kit receptor present on mast cells and stem cell factor (SCF, the ligand for ckit) expressed on tumor cells. Finally, we showed that targeting the SCF/cKit interaction with an anti-ckit antibody reduces the differentiation of mast cells and consequently reduces metastasis.

Conclusion: This is the first report to show that mast cells may play a critical role in remodeling not only the tumor microenvironment but also the metastatic niche to facilitate efficient metastasis through SCF/cKit interaction in breast cancer with arthritis.

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Increased migration and differentiation. (A through G) Increased differentiation of mast cells (MCs) from bone marrow (BM)-derived hematopoeitic precursors in arthritic versus nonarthritic mice with breast cancer (BC). Significant increase in MC count in (A) ± tumor-bearing SKG (*P < 0.05; **P < 001) and (C) arthritic C57BL/6 and PyV MT (*P < 0.05; **P < 001) mice (10 mice). (B, D) Representative images of MCs at 400× magnification. (E, F) Flow-cytometric analysis of percentage of cells expressing cKit receptor (CD117) on MCs. (G) Representative image of toluidine staining for MCs (1,000× magnification). (H, I) Increased migration of tumor cells toward MCs derived from arthritic tumor-bearing mice. Significant increase in migration of 4T1 and PyV MT cells toward the MCs from tumor-bearing arthritic mice (BMMCs) (**P < 001 and *P < 0.05). (H, I) Treatment of tumor cells with anti-SCF antibody or MCs with anti-c-Kit antibody decreased the migration of tumor cells toward mast cells. Pretreatment of 4T1 and PyV MT cells with anti-SCF antibody or adding anti-c-Kit antibody to the MCs in the lower chamber significantly decreased the migration of the tumor cells toward the MCs (*P < 0.05; **P < 0.01; ***P < 0.001).
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Figure 7: Increased migration and differentiation. (A through G) Increased differentiation of mast cells (MCs) from bone marrow (BM)-derived hematopoeitic precursors in arthritic versus nonarthritic mice with breast cancer (BC). Significant increase in MC count in (A) ± tumor-bearing SKG (*P < 0.05; **P < 001) and (C) arthritic C57BL/6 and PyV MT (*P < 0.05; **P < 001) mice (10 mice). (B, D) Representative images of MCs at 400× magnification. (E, F) Flow-cytometric analysis of percentage of cells expressing cKit receptor (CD117) on MCs. (G) Representative image of toluidine staining for MCs (1,000× magnification). (H, I) Increased migration of tumor cells toward MCs derived from arthritic tumor-bearing mice. Significant increase in migration of 4T1 and PyV MT cells toward the MCs from tumor-bearing arthritic mice (BMMCs) (**P < 001 and *P < 0.05). (H, I) Treatment of tumor cells with anti-SCF antibody or MCs with anti-c-Kit antibody decreased the migration of tumor cells toward mast cells. Pretreatment of 4T1 and PyV MT cells with anti-SCF antibody or adding anti-c-Kit antibody to the MCs in the lower chamber significantly decreased the migration of the tumor cells toward the MCs (*P < 0.05; **P < 0.01; ***P < 0.001).

Mentions: Because we observed increased MCs in the arthritic mice even in the absence of a tumor, we determined whether the differentiation of MCs from the bone marrow-derived precursors was affected by the arthritic milieu. We cultured 2 × 106 BM cells in the presence of IL-3 and SCF for 30 days to induce the precursor cells to differentiate into MCs. BM cells differentiate into MCs under selection with recombinant IL-3 and SCF [30]. We observed significant increase in the bone marrow-derived mast cells (BMMCs) in non-tumor-bearing arthritic SKG and C57BL/6+CII mice versus Balb/C with tumor or PyV MT mice (Figure 7A and 7C; Tables 1 and 2). Balb/C or C57BL/6 had no detectable levels of mast cells (Table 1 and 2). Numbers of MCs are represented in Figure 7A and 7C, respectively. Light-microscopic images of differentiated MCs from representative groups are shown in Figure 7B and 7D, respectively. Significant increase in MC differentiation was observed in tumor-bearing SKG and arthritic PyV MT mice as compared with tumor-bearing nonarthritic Balb/C mice and nonarthritic PyV MT mice (Figure 7A through D). Because differentiated mast cells should express the c-Kit receptor, we stained the cells with a c-Kit antibody (CD117) and analyzed with flow cytometry. A representative histogram is shown in Figure 7E and 7F. To confirm that the differentiated cells were indeed MCs, toluidine staining was conducted, and representative staining is shown in Figure 7G. Data suggest that the BM-derived precursor cells that are predestined to differentiate into MCs are significantly higher in the arthritic BM milieu before tumors are formed and that the tumors further enhance the differentiation (Tables 1 and 2). It can therefore be speculated that AA probably affects hematopoiesis and induces the production of MCs, thus creating a microenvironment appropriate for tumor cells to home and form metastases.


Arthritis augments breast cancer metastasis: role of mast cells and SCF/c-Kit signaling.

Das Roy L, Curry JM, Sahraei M, Besmer DM, Kidiyoor A, Gruber HE, Mukherjee P - Breast Cancer Res. (2013)

Increased migration and differentiation. (A through G) Increased differentiation of mast cells (MCs) from bone marrow (BM)-derived hematopoeitic precursors in arthritic versus nonarthritic mice with breast cancer (BC). Significant increase in MC count in (A) ± tumor-bearing SKG (*P < 0.05; **P < 001) and (C) arthritic C57BL/6 and PyV MT (*P < 0.05; **P < 001) mice (10 mice). (B, D) Representative images of MCs at 400× magnification. (E, F) Flow-cytometric analysis of percentage of cells expressing cKit receptor (CD117) on MCs. (G) Representative image of toluidine staining for MCs (1,000× magnification). (H, I) Increased migration of tumor cells toward MCs derived from arthritic tumor-bearing mice. Significant increase in migration of 4T1 and PyV MT cells toward the MCs from tumor-bearing arthritic mice (BMMCs) (**P < 001 and *P < 0.05). (H, I) Treatment of tumor cells with anti-SCF antibody or MCs with anti-c-Kit antibody decreased the migration of tumor cells toward mast cells. Pretreatment of 4T1 and PyV MT cells with anti-SCF antibody or adding anti-c-Kit antibody to the MCs in the lower chamber significantly decreased the migration of the tumor cells toward the MCs (*P < 0.05; **P < 0.01; ***P < 0.001).
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Figure 7: Increased migration and differentiation. (A through G) Increased differentiation of mast cells (MCs) from bone marrow (BM)-derived hematopoeitic precursors in arthritic versus nonarthritic mice with breast cancer (BC). Significant increase in MC count in (A) ± tumor-bearing SKG (*P < 0.05; **P < 001) and (C) arthritic C57BL/6 and PyV MT (*P < 0.05; **P < 001) mice (10 mice). (B, D) Representative images of MCs at 400× magnification. (E, F) Flow-cytometric analysis of percentage of cells expressing cKit receptor (CD117) on MCs. (G) Representative image of toluidine staining for MCs (1,000× magnification). (H, I) Increased migration of tumor cells toward MCs derived from arthritic tumor-bearing mice. Significant increase in migration of 4T1 and PyV MT cells toward the MCs from tumor-bearing arthritic mice (BMMCs) (**P < 001 and *P < 0.05). (H, I) Treatment of tumor cells with anti-SCF antibody or MCs with anti-c-Kit antibody decreased the migration of tumor cells toward mast cells. Pretreatment of 4T1 and PyV MT cells with anti-SCF antibody or adding anti-c-Kit antibody to the MCs in the lower chamber significantly decreased the migration of the tumor cells toward the MCs (*P < 0.05; **P < 0.01; ***P < 0.001).
Mentions: Because we observed increased MCs in the arthritic mice even in the absence of a tumor, we determined whether the differentiation of MCs from the bone marrow-derived precursors was affected by the arthritic milieu. We cultured 2 × 106 BM cells in the presence of IL-3 and SCF for 30 days to induce the precursor cells to differentiate into MCs. BM cells differentiate into MCs under selection with recombinant IL-3 and SCF [30]. We observed significant increase in the bone marrow-derived mast cells (BMMCs) in non-tumor-bearing arthritic SKG and C57BL/6+CII mice versus Balb/C with tumor or PyV MT mice (Figure 7A and 7C; Tables 1 and 2). Balb/C or C57BL/6 had no detectable levels of mast cells (Table 1 and 2). Numbers of MCs are represented in Figure 7A and 7C, respectively. Light-microscopic images of differentiated MCs from representative groups are shown in Figure 7B and 7D, respectively. Significant increase in MC differentiation was observed in tumor-bearing SKG and arthritic PyV MT mice as compared with tumor-bearing nonarthritic Balb/C mice and nonarthritic PyV MT mice (Figure 7A through D). Because differentiated mast cells should express the c-Kit receptor, we stained the cells with a c-Kit antibody (CD117) and analyzed with flow cytometry. A representative histogram is shown in Figure 7E and 7F. To confirm that the differentiated cells were indeed MCs, toluidine staining was conducted, and representative staining is shown in Figure 7G. Data suggest that the BM-derived precursor cells that are predestined to differentiate into MCs are significantly higher in the arthritic BM milieu before tumors are formed and that the tumors further enhance the differentiation (Tables 1 and 2). It can therefore be speculated that AA probably affects hematopoiesis and induces the production of MCs, thus creating a microenvironment appropriate for tumor cells to home and form metastases.

Bottom Line: We recently reported that mice with autoimmune arthritis have significantly increased incidence of bone and lung metastasis and decreased survival associated with breast cancer.This may primarily be due to the interaction of c-Kit receptor present on mast cells and stem cell factor (SCF, the ligand for ckit) expressed on tumor cells.Finally, we showed that targeting the SCF/cKit interaction with an anti-ckit antibody reduces the differentiation of mast cells and consequently reduces metastasis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Breast cancer remains the second leading cause of cancer-related deaths for women in the United States. Metastasis is regulated not only by intrinsic genetic changes in malignant cells, but also by the microenvironment, especially those associated with chronic inflammation. We recently reported that mice with autoimmune arthritis have significantly increased incidence of bone and lung metastasis and decreased survival associated with breast cancer. In this study, we evaluated the mechanism underlying the increased metastasis.

Methods: We used two mouse models; one that develops spontaneous autoimmune arthritis (SKG mice) injected with metastatic breast cancer cells (4T1), and another that develops spontaneous breast cancer (MMTV-PyV MT mice) injected with type II collagen to induce autoimmune arthritis. Mast cell levels and metastasis were monitored.

Results: First, we confirmed that breast tumor-bearing arthritic mice have a significantly higher incidence of bone and lung metastasis than do their nonarthritic counterparts. Next, we showed increased recruitment of mast cells within the primary tumor of arthritic mice, which facilitates metastasis. Next, we report that arthritic mice without any tumors have higher numbers of mast cells in the bones and lungs, which may be the underlying cause for the enhanced lung and bone metastases observed in the arthritic mice. Next, we showed that once the tumor cells populate the metastatic niches (bones and lungs), they further increase the mast cell population within the niche and assist in enhancing metastasis. This may primarily be due to the interaction of c-Kit receptor present on mast cells and stem cell factor (SCF, the ligand for ckit) expressed on tumor cells. Finally, we showed that targeting the SCF/cKit interaction with an anti-ckit antibody reduces the differentiation of mast cells and consequently reduces metastasis.

Conclusion: This is the first report to show that mast cells may play a critical role in remodeling not only the tumor microenvironment but also the metastatic niche to facilitate efficient metastasis through SCF/cKit interaction in breast cancer with arthritis.

Show MeSH
Related in: MedlinePlus