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Vav2 protein overexpression marks and may predict the aggressive subtype of ductal carcinoma in situ.

Jiang Y, Prabakaran I, Wan F, Mitra N, Furstenau DK, Hung RK, Cao S, Zhang PJ, Fraker DL, Guvakova MA - Biomark Res (2014)

Bottom Line: To date, there are no effective predictive biomarkers for identifying this subset with worse prognosis whose lesions are essentially indistinguishable histologically from those with favorable outcomes.Using a novel imaging-based method of tissue testing, we measured the relative expression levels of three candidate BM proteins specifically implicated in IBC progression - the insulin-like growth factor I receptor (IGF-IR), Ras-related protein 1 (Rap1), and Vav2 oncoprotein.Protein profiles were compared in 42 histologically normal mammary epithelial samples, 71 CIS (35 without/36 with invasion either on diagnostic biopsy or final surgical excision), and 98 IBC of known estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrine and Oncologic Surgery, Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA USA.

ABSTRACT

Background: A subset of patients with ductal carcinoma in situ (DCIS) will develop invasive breast cancer (IBC). To date, there are no effective predictive biomarkers for identifying this subset with worse prognosis whose lesions are essentially indistinguishable histologically from those with favorable outcomes. We hypothesized that measurable parameters that discriminate DCIS from DCIS with concurrent invasion may serve as diagnostic biomarkers (BM) of progressive cancer in situ (CIS).

Results: Using a novel imaging-based method of tissue testing, we measured the relative expression levels of three candidate BM proteins specifically implicated in IBC progression - the insulin-like growth factor I receptor (IGF-IR), Ras-related protein 1 (Rap1), and Vav2 oncoprotein. Protein profiles were compared in 42 histologically normal mammary epithelial samples, 71 CIS (35 without/36 with invasion either on diagnostic biopsy or final surgical excision), and 98 IBC of known estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status. The levels of the IGF-IR and Rap1 protein expression were significantly elevated in ER-positive (ER+/PR+/-/HER2 -) DCIS relative to normal epithelium (P <0.0001). The IGF-IR protein expression was also significantly up regulated in HER2-positive (ER+/-/PR+/-/HER2+) DCIS relative to normal epithelium (P = 0.0002). IGF-IR and Rap1 protein expression levels were similar among DCIS patients without or with concurrent invasion. Vav2 upregulation in DCIS relative to normal group was not associated with steroid hormone receptor and HER2 status, but was associated with the presence of concurrent invasion, including microinvasion (invasive foci of less than 1 mm). DCIS with high Vav2 were more than twice as likely to progress to invasive cancers as DCIS with low Vav2 (odds ratio, 2.42; 95% CI, 1.26-4-65; P =0.008). Furthermore, a receiver operating characteristic curve analysis revealed moderate ability of Vav2 protein expression measurements in DCIS to predict the existence of invasion concurrent with DCIS (area under the curve, 0.71; 95% CI, 0.59- 0.84).

Conclusions: Our novel findings hold promise for utilizing Vav2 protein as a predictive BM for differentiating progressive from non-progressive DCIS.

No MeSH data available.


Related in: MedlinePlus

Differential expression of IGF-IR, Rap1, and Vav2 in DCIS, DCIS with microinvasion, DCIS/LCIS with invasion >1 mm. (A) Representative monochrome images of Vav2 staining in normal mammary epithelium, DCIS, DCIS/T1mic, DCIS/IDC, and IDC. Original magnification (x200). Insets, enlarged images of epithelial cells. Arrows, suspected microinvasion. (B) Box plots for unaggregated measurements of IGF-IR, Rap1, and Vav2 in the groups of tissue: normal mammary epithelium, DCIS, DCIS/T1mic, and DCIS/IDC + LCIS/ILC. +, mean value. Horizontal lines, medians. Boxes, 25th and 75th percentile. Whiskers, 75th +1.5 x IQR and 25th -1.5 x IQR. Dots, group outliers. Y axis, protein levels in relative units. (C) The ROC curves for averaged measurements of IGF-IR, Rap1, and Vav2 protein expression.
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Fig3: Differential expression of IGF-IR, Rap1, and Vav2 in DCIS, DCIS with microinvasion, DCIS/LCIS with invasion >1 mm. (A) Representative monochrome images of Vav2 staining in normal mammary epithelium, DCIS, DCIS/T1mic, DCIS/IDC, and IDC. Original magnification (x200). Insets, enlarged images of epithelial cells. Arrows, suspected microinvasion. (B) Box plots for unaggregated measurements of IGF-IR, Rap1, and Vav2 in the groups of tissue: normal mammary epithelium, DCIS, DCIS/T1mic, and DCIS/IDC + LCIS/ILC. +, mean value. Horizontal lines, medians. Boxes, 25th and 75th percentile. Whiskers, 75th +1.5 x IQR and 25th -1.5 x IQR. Dots, group outliers. Y axis, protein levels in relative units. (C) The ROC curves for averaged measurements of IGF-IR, Rap1, and Vav2 protein expression.

Mentions: Because Vav2 was increased in IBC, but not in DCIS lesions, we hypothesized that change in Vav2 protein expression might be associated specifically with the onset of invasive potential in tumor cells, i.e. invasive tumor progression. If so, Vav2 protein up-regulation could be a predictor for the development of invasive potential in tumors without morphologic signs of invasion. To test this hypothesis, we stratified 71 samples of CIS into three histologically different groups with regard to invasion: 35 DCIS without concurrent invasion (DCIS), 11 DCIS associated with microinvasive (<0.1 cm) carcinoma (DCIS/T1mic) and 21DCIS/4LCIS associated with >0.1 cm areas of invasion (DCIS/IDC + LCIS/ILC). It is worth emphasizing that our cohort of DCIS samples are patient- matched cases: DCIS on CNB and subsequent excisions with DCIS without or with microinvasion. This cohort was selected on review of a total of 928 records of DCIS on CNB, with 19.7% of matched cases found to be with microinvasion on subsequent excision, but not on preceding biopsy. While T1mic was not necessarily present on slides that were cut freshly from diagnostic blocks for our analysis, T1mic was documented on CNB or subsequent excision pathology reports. This stringent criterion of selection of DCIS vs. DCIS/T1 mic was applied to stratify DCIS samples, as likely as feasible, into indolent (without invasion) and progressing (with microinvasion) lesions. More importantly, among those cases we only selected DCIS in CNB that involved at least 4 ducts in size to ensure adequate DCIS sampling to represent the disease process and avoid risk of exhausting DCIS tissue for future patient care use. As illustrated in Figure 3A, in normal tissues and DCIS, Vav2 expression was associated with the cell membrane. Vav2 protein was detected on the cell membrane and the cytoplasm in DCIS with invasion and in IDC itself. Remarkably, Vav2 levels in the DCIS group were similar to the normal group (P = 0.99), but were increased in DCIS /T1mic and further significantly increased in DCIS/IDC + LCIS/ILC (P = 0.03). Rather unexpectedly, compared to normal epithelium, significant increases in IGF-IR (P = 0.0025) and Rap1 (P = 0.007) were found in earliest proliferative lesions of DCIS without further changes in DCIS/T1mic and DCIS/IDC + LCIS/ILC (Figure 3B). The area under the ROC curve indicates low abilities of IGF-IR and Rap1 measurements to discriminate CIS subgroups (Figure 3C). In marked contrast, Vav2 measurements distinguished CIS with invasion from pure DCIS and normal cells (AUC, 0.71; 95% CI 0.59- 0.84). Moreover, patients that had high levels of Vav2 protein expression in DCIS were more than twice as likely to have concurrent invasion than those with low levels of Vav2 (OR, 2.42; 95% CI 1.26-4-65; P = 0.008).Figure 3


Vav2 protein overexpression marks and may predict the aggressive subtype of ductal carcinoma in situ.

Jiang Y, Prabakaran I, Wan F, Mitra N, Furstenau DK, Hung RK, Cao S, Zhang PJ, Fraker DL, Guvakova MA - Biomark Res (2014)

Differential expression of IGF-IR, Rap1, and Vav2 in DCIS, DCIS with microinvasion, DCIS/LCIS with invasion >1 mm. (A) Representative monochrome images of Vav2 staining in normal mammary epithelium, DCIS, DCIS/T1mic, DCIS/IDC, and IDC. Original magnification (x200). Insets, enlarged images of epithelial cells. Arrows, suspected microinvasion. (B) Box plots for unaggregated measurements of IGF-IR, Rap1, and Vav2 in the groups of tissue: normal mammary epithelium, DCIS, DCIS/T1mic, and DCIS/IDC + LCIS/ILC. +, mean value. Horizontal lines, medians. Boxes, 25th and 75th percentile. Whiskers, 75th +1.5 x IQR and 25th -1.5 x IQR. Dots, group outliers. Y axis, protein levels in relative units. (C) The ROC curves for averaged measurements of IGF-IR, Rap1, and Vav2 protein expression.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4362647&req=5

Fig3: Differential expression of IGF-IR, Rap1, and Vav2 in DCIS, DCIS with microinvasion, DCIS/LCIS with invasion >1 mm. (A) Representative monochrome images of Vav2 staining in normal mammary epithelium, DCIS, DCIS/T1mic, DCIS/IDC, and IDC. Original magnification (x200). Insets, enlarged images of epithelial cells. Arrows, suspected microinvasion. (B) Box plots for unaggregated measurements of IGF-IR, Rap1, and Vav2 in the groups of tissue: normal mammary epithelium, DCIS, DCIS/T1mic, and DCIS/IDC + LCIS/ILC. +, mean value. Horizontal lines, medians. Boxes, 25th and 75th percentile. Whiskers, 75th +1.5 x IQR and 25th -1.5 x IQR. Dots, group outliers. Y axis, protein levels in relative units. (C) The ROC curves for averaged measurements of IGF-IR, Rap1, and Vav2 protein expression.
Mentions: Because Vav2 was increased in IBC, but not in DCIS lesions, we hypothesized that change in Vav2 protein expression might be associated specifically with the onset of invasive potential in tumor cells, i.e. invasive tumor progression. If so, Vav2 protein up-regulation could be a predictor for the development of invasive potential in tumors without morphologic signs of invasion. To test this hypothesis, we stratified 71 samples of CIS into three histologically different groups with regard to invasion: 35 DCIS without concurrent invasion (DCIS), 11 DCIS associated with microinvasive (<0.1 cm) carcinoma (DCIS/T1mic) and 21DCIS/4LCIS associated with >0.1 cm areas of invasion (DCIS/IDC + LCIS/ILC). It is worth emphasizing that our cohort of DCIS samples are patient- matched cases: DCIS on CNB and subsequent excisions with DCIS without or with microinvasion. This cohort was selected on review of a total of 928 records of DCIS on CNB, with 19.7% of matched cases found to be with microinvasion on subsequent excision, but not on preceding biopsy. While T1mic was not necessarily present on slides that were cut freshly from diagnostic blocks for our analysis, T1mic was documented on CNB or subsequent excision pathology reports. This stringent criterion of selection of DCIS vs. DCIS/T1 mic was applied to stratify DCIS samples, as likely as feasible, into indolent (without invasion) and progressing (with microinvasion) lesions. More importantly, among those cases we only selected DCIS in CNB that involved at least 4 ducts in size to ensure adequate DCIS sampling to represent the disease process and avoid risk of exhausting DCIS tissue for future patient care use. As illustrated in Figure 3A, in normal tissues and DCIS, Vav2 expression was associated with the cell membrane. Vav2 protein was detected on the cell membrane and the cytoplasm in DCIS with invasion and in IDC itself. Remarkably, Vav2 levels in the DCIS group were similar to the normal group (P = 0.99), but were increased in DCIS /T1mic and further significantly increased in DCIS/IDC + LCIS/ILC (P = 0.03). Rather unexpectedly, compared to normal epithelium, significant increases in IGF-IR (P = 0.0025) and Rap1 (P = 0.007) were found in earliest proliferative lesions of DCIS without further changes in DCIS/T1mic and DCIS/IDC + LCIS/ILC (Figure 3B). The area under the ROC curve indicates low abilities of IGF-IR and Rap1 measurements to discriminate CIS subgroups (Figure 3C). In marked contrast, Vav2 measurements distinguished CIS with invasion from pure DCIS and normal cells (AUC, 0.71; 95% CI 0.59- 0.84). Moreover, patients that had high levels of Vav2 protein expression in DCIS were more than twice as likely to have concurrent invasion than those with low levels of Vav2 (OR, 2.42; 95% CI 1.26-4-65; P = 0.008).Figure 3

Bottom Line: To date, there are no effective predictive biomarkers for identifying this subset with worse prognosis whose lesions are essentially indistinguishable histologically from those with favorable outcomes.Using a novel imaging-based method of tissue testing, we measured the relative expression levels of three candidate BM proteins specifically implicated in IBC progression - the insulin-like growth factor I receptor (IGF-IR), Ras-related protein 1 (Rap1), and Vav2 oncoprotein.Protein profiles were compared in 42 histologically normal mammary epithelial samples, 71 CIS (35 without/36 with invasion either on diagnostic biopsy or final surgical excision), and 98 IBC of known estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status.

View Article: PubMed Central - PubMed

Affiliation: Division of Endocrine and Oncologic Surgery, Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA USA.

ABSTRACT

Background: A subset of patients with ductal carcinoma in situ (DCIS) will develop invasive breast cancer (IBC). To date, there are no effective predictive biomarkers for identifying this subset with worse prognosis whose lesions are essentially indistinguishable histologically from those with favorable outcomes. We hypothesized that measurable parameters that discriminate DCIS from DCIS with concurrent invasion may serve as diagnostic biomarkers (BM) of progressive cancer in situ (CIS).

Results: Using a novel imaging-based method of tissue testing, we measured the relative expression levels of three candidate BM proteins specifically implicated in IBC progression - the insulin-like growth factor I receptor (IGF-IR), Ras-related protein 1 (Rap1), and Vav2 oncoprotein. Protein profiles were compared in 42 histologically normal mammary epithelial samples, 71 CIS (35 without/36 with invasion either on diagnostic biopsy or final surgical excision), and 98 IBC of known estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status. The levels of the IGF-IR and Rap1 protein expression were significantly elevated in ER-positive (ER+/PR+/-/HER2 -) DCIS relative to normal epithelium (P <0.0001). The IGF-IR protein expression was also significantly up regulated in HER2-positive (ER+/-/PR+/-/HER2+) DCIS relative to normal epithelium (P = 0.0002). IGF-IR and Rap1 protein expression levels were similar among DCIS patients without or with concurrent invasion. Vav2 upregulation in DCIS relative to normal group was not associated with steroid hormone receptor and HER2 status, but was associated with the presence of concurrent invasion, including microinvasion (invasive foci of less than 1 mm). DCIS with high Vav2 were more than twice as likely to progress to invasive cancers as DCIS with low Vav2 (odds ratio, 2.42; 95% CI, 1.26-4-65; P =0.008). Furthermore, a receiver operating characteristic curve analysis revealed moderate ability of Vav2 protein expression measurements in DCIS to predict the existence of invasion concurrent with DCIS (area under the curve, 0.71; 95% CI, 0.59- 0.84).

Conclusions: Our novel findings hold promise for utilizing Vav2 protein as a predictive BM for differentiating progressive from non-progressive DCIS.

No MeSH data available.


Related in: MedlinePlus