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Identification of stromal ColXα1 and tumor-infiltrating lymphocytes as putative predictive markers of neoadjuvant therapy in estrogen receptor-positive/HER2-positive breast cancer.

Brodsky AS, Xiong J, Yang D, Schorl C, Fenton MA, Graves TA, Sikov WM, Resnick MB, Wang Y - BMC Cancer (2016)

Bottom Line: We performed gene expression profiling on pre-NAC+H tumor samples from responding (no or minimal residual disease at surgery) and non-responding patients.Increased expression of genes encoding for stromal collagens, including Col10A1, and reduced expression of immune-associated genes, reflecting lower levels of total tumor-infiltrating lymphocytes (TILs), were strongly associated with poor pathologic response.ROC analysis suggests strong specificity and sensitivity for this combination in predicting treatment response.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Rhode Island Hospital and Lifespan Medical Center, Warren Alpert Medical School of Brown University, Providence, USA. alex_brodsky@brown.edu.

ABSTRACT

Background: The influence of the tumor microenvironment and tumor-stromal interactions on the heterogeneity of response within breast cancer subtypes have just begun to be explored. This study focuses on patients with estrogen receptor-positive/human epidermal growth factor receptor 2-positive (ER+/HER2+) breast cancer receiving neoadjuvant chemotherapy and HER2-targeted therapy (NAC+H), and was designed to identify novel predictive biomarkers by combining gene expression analysis and immunohistochemistry with pathologic response.

Methods: We performed gene expression profiling on pre-NAC+H tumor samples from responding (no or minimal residual disease at surgery) and non-responding patients. Gene set enrichment analysis identified potentially relevant pathways, and immunohistochemical staining of pre-treatment biopsies was used to measure protein levels of those pathways, which were correlated with pathologic response in both univariate and multivariate analysis.

Results: Increased expression of genes encoding for stromal collagens, including Col10A1, and reduced expression of immune-associated genes, reflecting lower levels of total tumor-infiltrating lymphocytes (TILs), were strongly associated with poor pathologic response. Lower TILs in tumor biopsies correlated with reduced likelihood of achieving an optimal pathologic response, but increased expression of the Col10A1 gene product, colXα1, had greater predictive value than stromal abundance for poor response (OR = 18.9, p = 0.003), and the combination of increased colXα1 expression and low TILs was significantly associated with poor response in multivariate analysis. ROC analysis suggests strong specificity and sensitivity for this combination in predicting treatment response.

Conclusions: Increased expression of stromal colXα1 and low TILs correlate with poor pathologic response in ER+/HER2+ breast tumors. Further studies are needed to confirm their predictive value and impact on long-term outcomes, and to determine whether this collagen exerts a protective effect on the cancer cells or simply reflects other factors within the tumor microenvironment.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemistry of colXα1. a Representative colXα1 immunostaining in low- and high- colXα1 expressing ER+/HER2+ breast cancers. Two representative cases, one with no response, RCBIII, and strong colXα1 signal, score = 2, and one with good response, RCB0, and no colXα1 signal, score = 0, are shown. Arrows indicate regions with tumor cells. b RNA levels as determined by the microarray correlate with colXα1 IHC signal in 9 cases
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Fig3: Immunohistochemistry of colXα1. a Representative colXα1 immunostaining in low- and high- colXα1 expressing ER+/HER2+ breast cancers. Two representative cases, one with no response, RCBIII, and strong colXα1 signal, score = 2, and one with good response, RCB0, and no colXα1 signal, score = 0, are shown. Arrows indicate regions with tumor cells. b RNA levels as determined by the microarray correlate with colXα1 IHC signal in 9 cases

Mentions: To evaluate the findings from the gene expression data that collagens are significantly associated with pCR, we tested the usefulness of an anti-colXα1 monoclonal antibody to predict poor response and evaluated its relationship with other microenvironment metrics including the amount of tumor-associated stroma and TILs for its role in pCR. We performed IHC in 10 reduction mammoplasty cases to define the colXα1 expression pattern in normal breast tissue. In normal breast tissue, stain was negative for colXα1 except for occasional faint staining in a perivascular distribution pattern (data not shown). Among the 74 ER+/HER2+ cases in our study group, 50 pre-treatment needle biopsy samples had sufficient residual material (at least 1 cm tumor/stroma in a 12 gauge needle core) to allow evaluation with anti- colXα1 IHC. The overall response rate (pCR + RCB I) in this set was 36 % (18 of 50 patients) Table 1. Microenvironmental factors including decreased amount of stroma (P = 0.016) and higher levels of TIL (P < 0.001) were associated with good response in these 50 cases (Table 1). In tumor samples, immunostaining of colXα1 was observed as intense peri- and intra-tumoral distribution in some tumors in the RCBIII case (Also see 20× image in Additional file 6: Figure S2). A periductal/perivascular colXα1 staining pattern was frequently observed (Fig. 3). Increased colXα1 staining was strongly associated with a poor response by a chi-squared test (P < 0.001) (Table 1). The two cases with no stroma were scored as having negative colXα1 staining as no signal was observed.Fig. 3


Identification of stromal ColXα1 and tumor-infiltrating lymphocytes as putative predictive markers of neoadjuvant therapy in estrogen receptor-positive/HER2-positive breast cancer.

Brodsky AS, Xiong J, Yang D, Schorl C, Fenton MA, Graves TA, Sikov WM, Resnick MB, Wang Y - BMC Cancer (2016)

Immunohistochemistry of colXα1. a Representative colXα1 immunostaining in low- and high- colXα1 expressing ER+/HER2+ breast cancers. Two representative cases, one with no response, RCBIII, and strong colXα1 signal, score = 2, and one with good response, RCB0, and no colXα1 signal, score = 0, are shown. Arrows indicate regions with tumor cells. b RNA levels as determined by the microarray correlate with colXα1 IHC signal in 9 cases
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835834&req=5

Fig3: Immunohistochemistry of colXα1. a Representative colXα1 immunostaining in low- and high- colXα1 expressing ER+/HER2+ breast cancers. Two representative cases, one with no response, RCBIII, and strong colXα1 signal, score = 2, and one with good response, RCB0, and no colXα1 signal, score = 0, are shown. Arrows indicate regions with tumor cells. b RNA levels as determined by the microarray correlate with colXα1 IHC signal in 9 cases
Mentions: To evaluate the findings from the gene expression data that collagens are significantly associated with pCR, we tested the usefulness of an anti-colXα1 monoclonal antibody to predict poor response and evaluated its relationship with other microenvironment metrics including the amount of tumor-associated stroma and TILs for its role in pCR. We performed IHC in 10 reduction mammoplasty cases to define the colXα1 expression pattern in normal breast tissue. In normal breast tissue, stain was negative for colXα1 except for occasional faint staining in a perivascular distribution pattern (data not shown). Among the 74 ER+/HER2+ cases in our study group, 50 pre-treatment needle biopsy samples had sufficient residual material (at least 1 cm tumor/stroma in a 12 gauge needle core) to allow evaluation with anti- colXα1 IHC. The overall response rate (pCR + RCB I) in this set was 36 % (18 of 50 patients) Table 1. Microenvironmental factors including decreased amount of stroma (P = 0.016) and higher levels of TIL (P < 0.001) were associated with good response in these 50 cases (Table 1). In tumor samples, immunostaining of colXα1 was observed as intense peri- and intra-tumoral distribution in some tumors in the RCBIII case (Also see 20× image in Additional file 6: Figure S2). A periductal/perivascular colXα1 staining pattern was frequently observed (Fig. 3). Increased colXα1 staining was strongly associated with a poor response by a chi-squared test (P < 0.001) (Table 1). The two cases with no stroma were scored as having negative colXα1 staining as no signal was observed.Fig. 3

Bottom Line: We performed gene expression profiling on pre-NAC+H tumor samples from responding (no or minimal residual disease at surgery) and non-responding patients.Increased expression of genes encoding for stromal collagens, including Col10A1, and reduced expression of immune-associated genes, reflecting lower levels of total tumor-infiltrating lymphocytes (TILs), were strongly associated with poor pathologic response.ROC analysis suggests strong specificity and sensitivity for this combination in predicting treatment response.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Rhode Island Hospital and Lifespan Medical Center, Warren Alpert Medical School of Brown University, Providence, USA. alex_brodsky@brown.edu.

ABSTRACT

Background: The influence of the tumor microenvironment and tumor-stromal interactions on the heterogeneity of response within breast cancer subtypes have just begun to be explored. This study focuses on patients with estrogen receptor-positive/human epidermal growth factor receptor 2-positive (ER+/HER2+) breast cancer receiving neoadjuvant chemotherapy and HER2-targeted therapy (NAC+H), and was designed to identify novel predictive biomarkers by combining gene expression analysis and immunohistochemistry with pathologic response.

Methods: We performed gene expression profiling on pre-NAC+H tumor samples from responding (no or minimal residual disease at surgery) and non-responding patients. Gene set enrichment analysis identified potentially relevant pathways, and immunohistochemical staining of pre-treatment biopsies was used to measure protein levels of those pathways, which were correlated with pathologic response in both univariate and multivariate analysis.

Results: Increased expression of genes encoding for stromal collagens, including Col10A1, and reduced expression of immune-associated genes, reflecting lower levels of total tumor-infiltrating lymphocytes (TILs), were strongly associated with poor pathologic response. Lower TILs in tumor biopsies correlated with reduced likelihood of achieving an optimal pathologic response, but increased expression of the Col10A1 gene product, colXα1, had greater predictive value than stromal abundance for poor response (OR = 18.9, p = 0.003), and the combination of increased colXα1 expression and low TILs was significantly associated with poor response in multivariate analysis. ROC analysis suggests strong specificity and sensitivity for this combination in predicting treatment response.

Conclusions: Increased expression of stromal colXα1 and low TILs correlate with poor pathologic response in ER+/HER2+ breast tumors. Further studies are needed to confirm their predictive value and impact on long-term outcomes, and to determine whether this collagen exerts a protective effect on the cancer cells or simply reflects other factors within the tumor microenvironment.

No MeSH data available.


Related in: MedlinePlus