A Pilot Study Assessing the Potential Role of non-CD133 Colorectal Cancer Stem Cells as Biomarkers

Abstract

Introduction: Over 50% of patients with colorectal cancer (CRC) will progress and/or develop metastases. Biomarkers capable of predicting progression, risk stratification and therapeutic benefit are needed. Cancer stem cells are thought to be responsible for tumor initiation, dissemination and treatment failure. Therefore, we hypothesized that CRC cancer stem cell markers (CRCSC) will identify a group of patients at high risk for progression.Methods: Paraffin-embedded tissue cores of normal (n=8), and histopathologically well-defined primary (n= 30) and metastatic (n=10) CRC were arrayed in duplicate on tissue microarrays (TMAs). Expression profiles of non-CD133 CRCSC (CD29, CD44, ALDH1A1, ALDH1B1, EpCam, and CD166) were detected by immunohistochemistry and the association with clinicopathological data and patient outcomes was determined using standard statistical methodology. An independent pathologist, blinded to the clinical data scored the samples. Scoring included percent positive cells (0 to 4, 0 = <10%, 1 = 10 - 24%, 2 = 25 - 49%, 3 = 50 - 74%, 4 = 75 - 100%), and the intensity of positively stained cells (0 to 4; 0 = no staining, 1 = diminutive intensity, 2 = low intensity, 3 = intermediate intensity, 4 = high intensity). The pathologic score represents the sum of these two values, reported in this paper as a combined IHC staining score (CSS).Results: Of 30 patients 7 were AJCC stage IIA, 10 stage IIIB, 7 stage IIIC and 6 stage IV. Median follow-up was 113 months. DFI was 17 months. Median overall survival (OS) was not reached. Stage-specific OS was: II - not reached; III - not reached; IV - 11 months. In a univariate analysis, poor OS was associated with loss of CD29 expression; median OS, 32 months vs. not reached for CSS 3-7 vs. >7.5, respectively; p=0.052 comparing entire curves, after adjustment. In a Cox model analysis, loss of CD29 exhibited a trend toward association with survival (p=0.098) after adjusting for the effect of stage (p=0.0076). Greater expression of ALDH1A1 was associated with increasing stage (p=0.042 over stages 2, 3b, 3c, and 4) while loss of CD29 expression exhibited a trend toward being associated with stages 3 and 4 (p=0.08). Compared to normal colon tissue, primary tumors were associated with increased expression of ALDH1B1 (p=0.008). ALD1H1B1 expression level differed according to whether the tumor was moderately or poorly differentiated, well differentiated, or mucinous; the highest expression levels were associated with moderately or poorly differentiated tumors (p=0.011). Lymph node metastases were associated with a trend toward decreased expression of EpCAM (p = 0.06) when comparing 0 vs. 1 vs. 2+ positive lymph nodes, as was CD29 (p = 0.08) when comparing 0 vs. any positive lymph nodes. Compared to normal colon tissue metastatic colon cancers from different patients were associated with increased ALDH1B1 expression (p=0.001) whereas CD29 expression was higher in normal colonic tissue (p=0.014).Conclusion: CD29 may be associated with survival as well as clinical stage and...

Reduction in Membranous Immunohistochemical Staining for the Intracellular Domain of Epithelial Cell Adhesion Molecule Correlates with Poor Patient Outcome in Primary Colorectal Adenocarcinoma

“…After 59 months of follow-up, epcam-positive disseminated tumour cells were found to be significantly associated with overall survival and recurrence-free survival, and with a high density of peritumoural lymphatic vessels. In another study of 40 patients (30 primary, 10 metastatic), lymph node metastases were found to be associated with a trend toward decreased expression of epcam (p = 0.06) 27 .…”

The biological complexity of colorectal cancer: insights into biomarkers for early detection and personalized care

“…The literature data show that high ALDH1 expression is associated with high tumor grade and bad prognosis of disease [Langan et al 2012;Chen et al 2015].…”

“…ALCAM expression is very variable in CRC tissues; moreover, its overexpression has been associated with shortened OS and poor patient prognosis. The use of ALCAM to identify a CD44+/EpCAM+ subpopulation of cancer cells has also been suggested [Lugli et al 2010;Tachezy et al 2012;Langan et al 2012].…”

Isolation and identification of mesenchymal stem cells from human mastoid bone marrow

“…The characteristics of the isolated cells were first evaluated by flow cytometry using CD markers to identify potential stem cells. CD105 (endoglin) is a member of the transforming growth factor beta family, which has been linked with multi-lineage differentiation capacity [9], while CD29 (integrin beta 1) is responsible for mediating cell-to-cell adhesion and is involved in cell growth, differ- [10]. CD90 (Thy-1) is expressed in colony forming cells with high proliferative potential and is also used to characterize stem cell populations.…”