Association Of CXCR4 Expression Research Articles

Abstract 670: CXCR4 expression in tumor associated circulating cells of patients with pancreatic cancer is prognostic for progression and overall survival

Abstract Background: CXCR4 is a chemotaxis receptor implicated in the progression of Pancreatic Cancer (PC) via the activation and invasion of tumor cells. To better elucidate its role in the PC metastasis, we examined the role of CXCR4 expression on a variety of tumor associated circulating cells (TACs) [i.e. circulating tumor cells (CTCs), epithelial to mesenchymal cells (EMTs), and cancer associated macrophage-like cells (CAMLs)] as it relates to patient outcomes. A PC cell line PANC-1 was used to model the activation and expression pathway of CXCR4 in cells. Then blood samples were procured from 30 PC patients prior to the start of therapeutic intent to evaluate CXCR4 expression in TACs. CAMLs, EMTs, and CTCs were found in 93%, 63%, and 30% of PC patients respectively, with high CXCR4 expression found to significantly relate with higher TAC populations in circulation. Further, in a 2-year univariate analysis it was found that high expression of CXCR4 in CAMLs or EMTs was significantly related to both progression free survival (PFS) and overall survival (OS). These data suggest that CXCR4 expression in circulating cells in PC is strongly related to the progression and metastatic spread of the disease. Methods: PANC-1 cells were used to examine the expression and upregulation of CXCR4 by staining with an anti-CXCR4 antibody after stimulation with Isoproterenol. A prospective pilot study was initiated using anonymized peripheral blood samples from (n=30) PC patients, obtained in accordance with local IRB regulations with informed consent. TACs were isolated from the blood using a low-flow CellSieve Microfiltration system and stained for cytokeratin, CXCR4, and CD45. Wilcoxon ranked sum test and univariate analysis were used to analyze the association of CXCR4 expression on all TACs and to evaluate 24-month PFS/OS. Results: Within the PANC-1 cell line, we observed a median expression of CXCR4 intensity of roughly 210 after activation and upregulation with Isoproterenol. Within TACs in the circulation, higher numbers of CAMLs (p=0.017), EMTs (p=0.001), and CTCs (p<0.001) were all significantly related to cells with ≥210 CXCR4 expression. Further, higher expression of CXCR4 on CAMLs correlated with worse PFS (HR=4.0, 95%CI 1.5-10.5, p=0.012) and worse OS (HR=4.8, 95%CI 1.7-13.1, p=0.006), with higher CXCR4 expression in EMTs also correlated with worse PFS (HR=4.6, 95%CI 1.2-15.4, p=0.033) and worse OS (HR=5.4, 95%CI 1.5-19.2, p=0.021). Expression of CXCR4 in CTCs was not significant for PFS or OS. Conclusion: Expression of chemotaxis related receptor CXCR4 appears to have a strong relationship to higher numbers of TACs in circulation. This higher CXCR4 expression in CAMLs and EMTs also appears to correlate with rapid progression and death, indicating a relationship CXCR4 in the spread of PC and its potential use as a target for therapy. Citation Format: Kirby P. Gardner, Susan Tsai, Mohammed Aldakkak, Stephen Gironda, Cha-Mei Tang, Daniel L. Adams. CXCR4 expression in tumor associated circulating cells of patients with pancreatic cancer is prognostic for progression and overall survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 670.

Association of CXCR4 Expression and Clinical Outcome in Different Subsets of De Novo Acute Myeloid Leukemia Patients.

Acute myeloid leukemia is a heterogeneous group of diseases characterized by the uncontrolled proliferation of hematopoietic stem cells (HSCs) and progenitor cells with a reduced capacity to differentiate into mature cells. CXC chemokine receptor (CXCR4) and its ligand stromal derived factor-1 (SDF-1/CXCL12) are important players involved in cross-talk between leukemia cells and the bone marrow (BM) microenvironment. The aim to study the association between the immunohistochemical CXCR4 expression and the clinical outcome of AML in adult Egyptian patients. Fifty-eight patients suffering from AML were recruited for this study, with an age range from 18 to 60 years and presenting from January 2013 to March 2017. All patients were subjected to complete blood count, BM aspiration, immunophenotyping, BM trephine biopsy, immunohistochemical staining with CXCR4 McAb and cytogenetics when feasible. CXCR4 was widely expressed (55.2%) among the studied patients. There was a significant relationship between CXCR4 and patients' outcomes. Fifteen (71.4%) patients who died were CXCR4 positive. The estimated mean time until death among CXCR4 negative cases was 37.6 ± 4.04 months which was longer than that of CXCR4 positive cases who had mean of 20.04 ± 4.9 months p = 0.016. The risk for death among CXCR4 positive cases was higher than CXCR4 negative cases with hazard ratio (HR) = 2.147 (p = 0.048). These results suggest that CXCR4 was expressed in a subset of AML patients and was associated with poor prognosis. CXCR4 expression appears to be an independent prognostic factor for survival in a heterogeneous group of AML patients.

Abstract 1126: Association of CXCR4 expression with lymph node metastasis in distal cholangiocarcinoma

Abstract BACKGROUND: The standard treatment of resectable distal bile duct cancer is pancreatoduodenectomy. However, prognosis is still poor because of a high rate of lymph node (LN) metastasis, underlining the need to further clarify the mechanism of LN metastasis. Recently accumulated evidence has shown that cancer cells can take advantage of the homeostatic mechanism of lymphocyte homing to metastasize to LNs. The CXCL12-CXCR4 axis modulates lymphocyte homing by inducing continuous migration of dendritic cells and T cells that are expressing chemokine receptor CXCR4 to LNs where chemokine CXCL12 is homeostatically secreted. OBJECTIVE: The aim of this study is to elucidate whether distal bile duct cancer cells express CXCR4 and investigate a possible association with LN metastasis. METHODS: This study enrolled 32 patients with resectable distal bile duct cancer including 19 patients with LN metastasis and 13 without. All patients underwent pancreatoduodenectomy at our department between 2002 and 2008 and have been followed up (Median observation period: 50.5 (4-184) months). Histopathologically, 27 cases were tubular adenocarcinoma, and 5 cases were other types of cancer. Immunostaining with resected specimens was undertaken to examine the expression of CXCR4 in cancer cells and determine whether its expression level is associated with lymph node metastasis. When 10% and more of tumor cells were stained, specimens were diagnosed as positive for CXCR4. The result of staining level was visually assessed. RESULTS: Cancer cells were stained heterogeneously by antibody for CXCR4. Normal cells of the bile duct were not stained, while damaged or desquamated cells were stained. With regard to the staining pattern, the cytoplasm and membrane were stained but the nucleus was not stained. 19 out of 32 cases were positive for CXCR4 expression (59.4%). 15 of 19 cases with LN metastasis were positive whereas 4 of 13 cases without LN metastasis were positive (p=0.0107). CXCR4 expression on cancer cells increased with advancing cancer cell undifferentiation in tubular adenocarcinoma; cancer cells were positive for CXCR4 in 0 of 5 (0%), 6 of 16 (37.5%), and 6 of 6 (100%) cases in tubular 1, tubular 2, and tubular 3 adenocarcinoma, respectively. CONCLUSIONS: Our results indicated that the expression of CXCR4 in cancer cells may be associated with LN metastasis in distal bile duct cancer. Citation Format: Yohsuke Yagawa, Atsushi Aruga, Nobuhiro Takeshita, Ryota Higuchi, Takehisa Yazawa, Keishi Tanigawa, Masakazu Yamamoto. Association of CXCR4 expression with lymph node metastasis in distal cholangiocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1126.

Association of CXCR4 expression with coronary collateralization in patients with chronic total coronary occlusion: A nested case–control study

ObjectiveCXCR4 signaling contributes to the development and progression of neovascularization. The objective of this study was to investigate whether CXCR4 expression in peripheral CD34+ cells associated with the coronary collateralization (CC) in patients with chronic total coronary occlusion (CTO). Methods and resultsWe measured CXCR4 expression in peripheral CD34+ cells and assessed its relation with CC in a nested case–control study including 78 cases and 78 matched controls aged 38–69years, assessed in January 2011 to December 2012 and with at least 1year of follow-up before the index date. Cases were defined as good coronary collateralization (GCC) according to the Rentrop scoring system (Rentrop score of 2 or 3); for each case, one age-matched control with poor coronary collateralization (PCC) (Rentrop score 0 or 1) was randomly selected from the study participants. Demographic, biochemical, and angiographic variables were collected. In multivariate analysis, the OR (95% CI) of CXCR4 expression was 0.018 (0.017 to 0.020) in patients with GCC versus PCC. Independent effect of CXCR4 expression on CC was (OR 0.012, 95% CI 0.010–0.014) when adjusted for other variables. A nonlinear relationship between CXCR4 expression and CC was observed. The CC degree increased when CXCR4 expression exceeded the turning point (30%) (OR 0.025, 95% CI 0.022–0.028; p<0.001). When the CXCR4 expression exceeded 75%, increased CXCR4 level could not promoted CC (OR 0.000, 95% CI 0.008–0.007; p=0.974). ConclusionIncreased CXCR4 level in peripheral CD34+ cells was associated with GCC in patients with CTO.

CXCR4 Expression in Gastric Cancer and Bone Marrow: Association with Hypoxia-Regulated Indices, Disseminated Tumor Cells, and Patients Survival

Aim. The analysis of the association of CXCR4 expression in gastric cancer (GC) and bone marrow (BM) with clinical characteristics. Patients and Methods. 65 patients with GC were investigated. Immunohistochemistry, immunocytochemistry, NMR-spectroscopy, and zymography were used. Results. CXCR4 was expressed in 78.5% of GC specimens and correlated with tumor hypoxia (P&lt;0.05), VEGF expression (P&lt;0.01), and gelatinases activity (P&lt;0.05). CXCR4-positive cells in GC were detected in 80% of patients with disseminated tumor cells (DTCs). Overall survival (OS) of patients with CXCR4-positive tumors was poorer than that of patients with CXCR4-negative tumors (P=0.037). The CXCR4-positive cells in BM were found in 46% of all patients and in 56% of patients with DTCs. CXCR4 expression in BM was not associated with OS. Risk of unfavourable outcome is increased in patients with CXCR4-positive tumors (P&lt;0.05). CXCR4 expression in BM was positively associated with DTCs, especially in patients with M0 category. Risk of unfavourable outcome is increased in patients with M0 category and with both CXCR4-positive BM and DTCs (P=0.03). Conclusions. CXCR4 expression in tumor was positively correlated with hypoxia level and VEGF expression in tumor as well as OS. CXCR4 expression in BM is associated with DTCs.

The association of CXCR4 expression with prognosis and clinicopathological indicators in colorectal carcinoma patients: a meta‐analysis

The clinical relevance of expression of chemokine receptor 4 (CXCR4) in colorectal carcinoma (CRC) remains controversial; our aim was to identify the precise relationship of CXCR4 to prognosis and clinicopathological features. A meta-analysis was performed. Original data included the hazard ratios (HRs) of recurrence-free survival (RFS), overall survival (OS) and odds ratio (OR) in CRC patients. We pooled HR/OR with 95% confidence intervals (CIs) to estimate the hazard. A total of 20 published studies (including 2253 patients) were eligible. RFS and OS were related significantly to CXCR4 expression, with HRs 1.62 (95% CI 1.24-2.11; P < 0.0001) and 1.68 (95% CI 1.31-2.14; P < 0.0001), respectively. In addition, a significant association was revealed between positive CXCR4 expression and age (less than median age: OR 0.78, 95% CI 0.62-0.98; P = 0.03), stage (I and II: OR 0.46, 95% CI 0.32-0.66; P < 0.0001), grade (well/moderately differentiated: OR 0.74, 95% CI 0.56-0.98; P = 0.04), location (colon: OR: 0.73, 95% CI 0.57-0.95; P = 0.02), lymph node invasion (present: OR2.14, 95% CI 1.36-3.37; P = 0.001),and distant metastasis (present: OR 2.40; 95% CI 1.36-4.23; P = 0.003). Heterogeneity was observed among the included studies with regard to stage (I(2) = 58 %), lymph node invasiveness (I(2) = 74%) and distant metastasis (I(2) = 56%). No publication bias was observed. Chemokine receptor 4 expression indicates poorer prognosis in older patients and advanced stage or poor differentiation in CRC, and also serves as an indicator of lymph node and distal organ metastasis. Surprisingly, high CXCR4 expression may indicate that the location of the tumour is the rectum. Thus, CXCR4 could help to predict outcome and guide clinical therapy.

Evaluation of the association of CXCR4 expression with HER2 status or mTOR expression levels

e22192 Introduction: CXCR4 is a chemokine receptor that is linked to breast cancer metastasis. The putative mechanism involves activation of the HER-2/PI3K/PTEN/Akt/mammalian target of rapamycin (mTOR) pathway, resulting in enhanced synthesis of downstream proteins such as CXCR4 and eukaryotic initiation factor 4E (eIF4E). Overexpression of eIF4E has been shown to be an independent predictor of breast cancer outcome. Because HER-2/mTOR/CXCR4 pathway has mainly been observed in vitro, it deserves scrutiny to determine whether it is also seen in human breast cancers. We hypothesize that HER-2/mTOR axis is not significantly associated with CXCR4 regulation in human breast cancers. Method: 107 stages 1 - 3 breast cancers from a prospective breast cancer database were analyzed. Samples were quantified for mTOR, CXCR4, and eIF4E levels by Western Blot. mTOR, and eIF4E levels were quantified as fold-over benign breast tissues and CXCR4 levels were quantified as fold-over Hela cells. CXCR4 overexpression was defined as low (&lt; 6.0 fold) or high (≥ 6.0 fold), and eIF4E expression level was separated into tertiles. Primary end points were cancer recurrence and death. Statistical analyses performed were independent student t-test, Spearman correlation, Kaplan-Meier survival curves, log-rank test, and Cox proportional hazard model. Results: All 59 HER-2 (-) and 48 HER-2 (+) breast cancer specimens overexpressed CXCR4 (mean=6.18 ±4.4), mTOR (mean=1.24 ±1.58), and eIF4E (mean=9.53 ±6.42). Unlike in vitro data, there was no difference in mean CXCR4 expression level between HER-2 (-) and HER-2 (+) tumors (mean=6.55, 5.72, respectively) (p=0.33). While mTOR correlated significantly with eIF4E level (p=0.0006, r = 0.34), it did not correlate with CXCR4 level (p=0.713, r=0.036). Only nodal status (p=0.04) and eIF4E level (p=0.03) were independent predictors of breast cancer death. Conclusions: We demonstrate that CXCR4 regulation in human breast cancers is more complex than previously thought. There was no correlation between mTOR and CXCR4 levels, nor was there a difference in mean CXCR4 overexpression levels between HER-2 (+) and HER-2 (-) tumors. Pathways regulating CXCR4 expression warrant further investigation. No significant financial relationships to disclose.

Nuclear expression of CXCR4 in tumor cells of non–small cell lung cancer is correlated with lymph node metastasis

The stromal-derived factor 1alpha (CXCL12)/chemokine receptor CXCR4 system plays an important role in the metastatic process of a variety of cancers, with CXCR4 frequently expressed by tumor cells homing to CXCL12-rich compartments. The current study evaluated a possible association of CXCR4 expression with lymph node metastasis in primary non-small cell lung cancer. CXCR4 expression levels were evaluated using immunohistology in 46 non-small cell lung cancer specimens of patients without or with lymph node involvement (N0 = 24, N1/N2/N3 = 22). Evaluation of immunostaining was performed semiquantitatively by visual assessment. Statistical analyses with multiple testing adjustments for confirmatory comparisons were performed to assess relevant parameters associated with lymph node metastases. In all samples of non-small cell lung cancer, tumor cells stained positively for cytoplasmic CXCR4. The intensity of the CXCR4 staining varied considerably between specimens: 2 (4%) tumors demonstrated weak cytoplasmic CXCR4, 22 (48%) intermediate, and 22 (48%) strong staining. Membranous staining was absent; however, nuclear staining of CXCR4 was observed in 5 non-small cell lung cancer samples. Statistical analyses of the association between presence of lymph node metastases and CXCR4 expression levels revealed that cytoplasmic CXCR4 expression was not associated with the presence of lymph node metastases. However, nuclear CXCR4 was significantly correlated with increasing lymph node stage (P = .008), linear-to-linear association. The association between aberrant expression of CXCR4 in the nucleus of non-small cell lung cancer and metastasis to lymph nodes points toward a potential tumor metastasis promoting function of nuclear CXCR4.