Alexandra Sebastianelli

The RUNX1 transcription factor is expressed in serous epithelial ovarian carcinoma and contributes to cell proliferation, migration and invasion

Previously, we have identified the RUNX1 gene as hypomethylated and overexpressed in post-chemotherapy (CT) primary cultures derived from epithelial ovarian cancer (EOC) patients, when compared with primary cultures derived from matched primary (prior to CT) tumors. Here we show that RUNX1 displays a trend of hypomethylation, although not significant, in omental metastases compared with primary EOC tumors. Surprisingly, RUNX1 displayed significantly higher expression not only in metastatic tissue, but also in high-grade primary tumors and even in low malignant potential tumors. The RUNX1 expression levels were almost identical in primary tumors and omental metastases, suggesting that RUNX1 hypomethylation might have a limited impact on its overexpression in advanced (metastatic) stage of the disease. Knockdown of the RUNX1 expression in EOC cells led to sharp decrease of cell proliferation and induced G1 cell cycle arrest. Moreover, RUNX1 suppression significantly inhibited EOC cell migration and invasion. Gene expression profiling and consecutive network and pathway analyses confirmed these findings, as numerous genes and pathways known previously to be implicated in ovarian tumorigenesis, including EOC tumor invasion and metastasis, were found to be downregulated upon RUNX1 suppression, while a number of pro-apoptotic genes and some EOC tumor suppressor genes were induced. Taken together, our data are indicative for a strong oncogenic potential of the RUNX1 gene in EOC progression and suggest that RUNX1 might be a novel EOC therapeutic target. Further studies are needed to more completely elucidate the functional implications of RUNX1 and other members of the RUNX gene family in ovarian tumorigenesis.

Simple vaginal trachelectomy in early-stage low-risk cervical cancer: a pilot study of 16 cases and review of the literature.

Objective This study aimed to evaluate the feasibility of simple vaginal trachelectomy and node assessment in patients with low-risk early-stage cervical cancer (<2 cm). Methods From May 2007 to November 2012, 16 women with low-risk small-volume cervical cancer underwent a simple vaginal trachelectomy preceded by laparoscopic sentinel node mapping plus or minus pelvic node dissection. Data were collected prospectively in a computerized database. Descriptive statistics and Kaplan-Meyer estimate were used for analysis. Results Patients’ median age was 30 years and 12 (75%) were nulliparous. Six had a diagnostic cone, 6 had a loop electrocautery excision procedure, 3 had cervical biopsies, and 1 had polyp excision. All patients underwent a preoperative pelvic magnetic resonance imaging. Four patients had stage IA1 with lymph vascular space invasion (LSVI), 6 IA2, and 6 IB1. Ten (62.5%) had squamous lesions, 7 had adenocarcinoma. LVSI was present in 4 cases, suspicious in 2, and absent in 10. There were 2 surgical complications: a trocar site hematoma and a vaginal laceration. The median OR time was 150 minutes (range, 120–180 minutes) and median blood loss was 50 mL (range, 50–150 mL). On final pathology, lymph nodes were negative in all patients. Thirteen (81%) patients had either no residual disease (6) or residual dysplasia only (7) in the trachelectomy specimen. Margins were negative in all cases. With a median follow-up of 27 months (range, 1–65 months), there have been no recurrences. The recurrence-free survival at 24 months is 100%. Eight patients have conceived: 3 were term deliveries and 4 are ongoing. Conclusions Simple trachelectomy and nodes seems to be a safe alternative in well-selected patients with early-stage low-risk cervical cancer. Our data will need to be confirmed in larger series.

Predictors of non-sentinel lymph node (non-SLN) metastasis in patients with sentinel lymph node (SLN) metastasis in endometrial cancer

OBJECTIVES The aim of this study was to determine the risk of metastasis in the remaining non-SLNs when the SLN is positive and to identify the factors that can predict this risk. METHODS We reviewed all patients who underwent primary surgery for endometrial carcinoma with lymphadenectomy and SLN mapping (November 2010-November 2013) in our center. SLNs were ultra-staged on final pathology. RESULTS A total of 268 patients were included. Overall, 43/268 patients (16%) were found to have SLN metastasis: macro-metastasis in 24 patients, micro-metastasis in 7 and ITC in 12. Non-SLN metastases were found in 15 of the 43 patients (34.8%) with positive SLN. Size of the SLN metastasis was the only factor associated with an increased likelihood of non-SLN metastasis (p=0.005). When the size of the SLN metastasis was ≤2mm, the risk of having another positive lymph node was only 5%, conversely, when the size of the SLN metastasis was >2mm, the risk of having another positive lymph node was 60.8% (p<0.0001). Histologic type, grade, depth of myometrial invasion, LVSI, cervical stromal invasion and CA-125 were not predictive. CONCLUSION When the SLN is positive, the risk of metastasis in the remaining non-SLNs was 34.8%. Size of the metastasis within the SLN was the only factor that could predict the risk of non-SLN metastasis; 2mm seems to be the cutoff size below which the risk of non-SLN metastasis is low.

BCAT1 expression associates with ovarian cancer progression: possible implications in altered disease metabolism

Previously, we have identified the branched chain amino-acid transaminase 1 (BCAT1) gene as notably hypomethylated in low-malignant potential (LMP) and high-grade (HG) serous epithelial ovarian tumors, compared to normal ovarian tissues. Here we show that BCAT1 is strongly overexpressed in both LMP and HG serous epithelial ovarian tumors, which probably correlates with its hypomethylated status. Knockdown of the BCAT1 expression in epithelial ovarian cancer (EOC) cells led to sharp decrease of cell proliferation, migration and invasion and inhibited cell cycle progression. BCAT1 silencing was associated with the suppression of numerous genes and pathways known previously to be implicated in ovarian tumorigenesis, and the induction of some tumor suppressor genes (TSGs). Moreover, BCAT1 suppression resulted in downregulation of numerous genes implicated in lipid production and protein synthesis, suggesting its important role in controlling EOC metabolism. Further metabolomic analyses were indicative for significant depletion of most amino acids and different phospho- and sphingolipids following BCAT1 knockdown. Finally, BCAT1 suppression led to significantly prolonged survival time in xenograft model of advanced peritoneal EOC. Taken together, our findings provide new insights about the functional role of BCAT1 in ovarian carcinogenesis and identify this transaminase as a novel EOC biomarker and putative EOC therapeutic target.

Preoperative CA 125 Tumour Marker in Endometrial Cancer: Correlation With Advanced Stage Disease

OBJECTIVE To evaluate if a preoperative serum CA 125 level>or=35 kU/L in patients with endometrial cancer correlates with a surgical stage III or IV and poor histopathological prognostic factors. METHODS We conducted a retrospective cohort study of 254 patients who underwent hysterectomy and full staging for endometrial cancer. Preoperative serum CA 125 was available for each patient as well as complete clinical and histopathological data. Chi-square, Fisher exact test, multivariate logistic regression, and receiver operating characteristic curve curves were used for statistical analysis. RESULTS A total of 186 (73%) patients had stage I or II disease and 68 (27%) had stage III or IV disease. A statistically higher number of patients from the stage III or IV group had a serum CA 125 level>or=35 kU/L (58%) compared with the stage I or II group (16%) (OR 7.44; P<0.001). There was no correlation between serum CA 125 level and histological subtype. Patients with stage I or II disease and serum CA 125>or=35 kU/L (46%) had significantly more frequent deep myometrial invasion (>50%) than did those with serum CA 125<35 kU/L (18%) (OR 3.68; P=0.006). CONCLUSION Assay of the preoperative serum CA 125 level is a very simple test to detect patients with more advanced stage endometrial adenocarcinoma. Its routine use could help triage high risk patients preoperatively.

Inhibition of RUNX2 Transcriptional Activity Blocks the Proliferation, Migration and Invasion of Epithelial Ovarian Carcinoma Cells

Previously, we have identified the RUNX2 gene as hypomethylated and overexpressed in post-chemotherapy (CT) primary cultures derived from serous epithelial ovarian cancer (EOC) patients, when compared to primary cultures derived from matched primary (prior to CT) tumors. However, we found no differences in the RUNX2 methylation in primary EOC tumors and EOC omental metastases, suggesting that DNA methylation-based epigenetic mechanisms have no impact on RUNX2 expression in advanced (metastatic) stage of the disease. Moreover, RUNX2 displayed significantly higher expression not only in metastatic tissue, but also in high-grade primary tumors and even in low malignant potential tumors. Knockdown of the RUNX2 expression in EOC cells led to a sharp decrease of cell proliferation and significantly inhibited EOC cell migration and invasion. Gene expression profiling and consecutive network and pathway analyses confirmed these findings, as various genes and pathways known previously to be implicated in ovarian tumorigenesis, including EOC tumor invasion and metastasis, were found to be downregulated upon RUNX2 suppression, while a number of pro-apoptotic genes and some EOC tumor suppressor genes were induced. Taken together, our data are indicative for a strong oncogenic potential of the RUNX2 gene in serous EOC progression and suggest that RUNX2 might be a novel EOC therapeutic target. Further studies are needed to more completely elucidate the functional implications of RUNX2 and other members of the RUNX gene family in ovarian tumorigenesis.

Isolated tumor cells identified by sentinel lymph node mapping in endometrial cancer: Does adjuvant treatment matter?

OBJECTIVE To evaluate the outcome and the role of adjuvant treatment in the management of patients with endometrial cancer and isolated tumor cells (ITCs) identified by SLN mapping. METHODS This single center study identified all patients undergoing hysterectomy, salpingo-oophorectomy, lymphadenectomy and SLN mapping for endometrial cancer between November 2010 and December 2015. Data was prospectively collected. Progression-free survival was analyzed according to the Kaplan-Meier method and compared using the log-rank test. RESULTS A total of 519 patients were included. Overall, 85 patients (16.4%) were found to have SLN metastases of which 43 (51%) were macrometastasis, 11 (13%) micrometastasis (MM) and 31 (36%) ITC. Eleven (35%) of patients with ITCs received adjuvant chemotherapy±whole pelvic radiation therapy (WPRT), 10 (32%) received WPRT, and 10 (32%) received either no adjuvant treatment or vault brachytherapy (VBT) only. ITC patients received significantly less chemotherapy (p=0.0001) and WPRT (p=0.007) compared to patients with macrometastasis. Of note, ITC were not considered node positive in our study. With a median follow-up of 29months (range: 0-67), the progression free survival (PFS) at 3-years for the ITC patients was 95.5%, similar to node negative (87.6%) and micrometastasis patients (85.5%), but statistically better than patients with macrometastasis (58.5%) (p=0.0012). Only 1/31 patient with ITC recurred (IB, 7cm carcinosarcoma) despite adjuvant treatments. None of the ITC patients with endometrioid histology recurred (0/28) and none of the ITC patients who did not receive adjuvant treatment or VBT recurred (0/10). CONCLUSIONS Patients with endometrial cancer found to have SLN ITCs have an excellent outcome. The use of adjuvant treatment should be tailored to uterine factors and histology and not solely based on the presence of ITCs. Patients with ITCs and otherwise low-risk uterine disease probably derive little benefit from receiving additional treatments. More studies are needed to confirm our results.

Performance of sentinel lymph node (SLN) mapping in high-risk endometrial cancer

OBJECTIVE While the accuracy of the SLN procedure has been validated in patients with low risk EC, its relevance for high-risk EC remains debated. The aim of this study was to evaluate the accuracy of SLN mapping in patients with high-risk EC. STUDY METHOD We reviewed all patients with high risk EC (grade 3 endometrioid, serous, carcinosarcoma, clear cell and undifferentiated) who underwent primary surgery with SLN mapping followed by pelvic +/- paraaortic lymphadenectomy, between November 2010 and November 2016. RESULTS Among 128 patients who underwent SLN mapping followed by a pelvic lymph node dissection, 41 (32%) had a positive pelvic lymph node. Overall, 48.8% of patients underwent paraaortic node sampling (62/128). Paraaortic lymph node metastasis was identified in 17.7% of patients in whom a para-aortic lymph node dissection was performed (11/62), and all had positive pelvic lymph nodes as well. Successful SLN mapping occurred in 115/128 (89,8%) patients, with a bilateral detection rate at 63.2% (81/128). Positive SLNs were identified in 30.4% of patients (39/128) including 7 isolated tumor cells (ITC), 4 micrometastasis and 28 macrometastasis. When the SLNs were detected bilaterally, only one false negative case occurred, providing a sensitivity and negative predictive value of 95.8% and 98.2% respectively. CONCLUSION Accurate surgical staging is an important prognostic predictor of survival in patients with endometrial cancer. Given the high sensitivity and high negative predictive value found in our study, we believe that the use of SLN mapping appears to be an appropriate staging procedure in high-risk endometrial cancer.

The mannose receptor LY75 (DEC205/CD205) modulates cellular phenotype and metastatic potential of ovarian cancer cells

The molecular basis of epithelial ovarian cancer (EOC) dissemination is still poorly understood. Previously, we identified the mannose receptor LY75 gene as hypomethylated in high-grade (HG) serous EOC tumors, compared to normal ovarian tissues. LY75 represents endocytic receptor expressed on dendritic cells and so far, has been primarily studied for its role in antigen processing and presentation. Here we demonstrate that LY75 is overexpressed in advanced EOC and that LY75 suppression induces mesenchymal-to-epithelial transition (MET) in EOC cell lines with mesenchymal morphology (SKOV3 and TOV112), accompanied by reduction of their migratory and invasive capacity in vitro and enhanced tumor cell colonization and metastatic growth in vivo. LY75 knockdown in SKOV3 cells also resulted in predominant upregulation of functional pathways implicated in cell proliferation and metabolism, while pathways associated with cell signaling and adhesion, complement activation and immune response were mostly suppressed. Moreover, LY75 suppression had an opposite effect on EOC cell lines with epithelial phenotype (A2780s and OV2008), by directing epithelial-to-mesenchymal transition (EMT) associated with reduced capacity for in vivo EOC cell colonization, as similar/identical signaling pathways were reversely regulated, when compared to mesenchymal LY75 knockdown EOC cells. To our knowledge, this is the first report of a gene displaying such pleiotropic effects in sustaining the cellular phenotype of EOC cells and points to novel functions of this receptor in modulating EOC dissemination. Our data also support previous findings regarding the superior capacity of epithelial cancer cells in metastatic colonization of distant sites, compared to cancer cells with mesenchymal-like morphology.

Simple Vaginal Trachelectomy: A Valuable Fertility-Preserving Option in Early-Stage Cervical Cancer

Objective Radical trachelectomy is a valid alternative for the treatment of early-stage cervical cancer in young women who wish to preserve fertility potential. Recent data indicate that even less radical surgery could be performed in low-risk cases. The objective of our study was to evaluate the safety of simple vaginal trachelectomy and node assessment in patients with low-risk, early-stage cervical cancer (<2 cm). Methods From May 2007 to July 2016, 35 women underwent a simple vaginal trachelectomy with laparoscopic sentinel lymph node mapping + pelvic node dissection. Data were collected prospectively in a computerized database. Descriptive statistics and Kaplan-Meier estimate were used for analysis. Results Patients’ median age was 29 years, and 24 (69%) were nulliparous. Eight had stage IA1 with lymphovascular space invasion, 9 a stage IA2, and 18 a stage IB1. Nineteen (54%) had squamous histology, 13 (37%) had adenocarcinoma, and 3 had other histologic findings. The median operating room time was 148 minutes (90–240 minutes), and median blood loss was 50 mL (25–200 mL). On final pathology, lymph nodes were negative in all patients, except 2 cases with isolated tumor cells. Twenty-two patients (63%) had either no residual disease in the trachelectomy specimen (n = 15) or residual dysplasia only (n = 7). With a median follow-up of 42 months (1–100 months), 1 local recurrence occurred treated initially with chemoradiation and then a pelvic exenteration. The recurrence-free survival at 48 months is 96.7%. There were 25 pregnancies: 5 (20%) ended in the first trimester, 2 delivered prematurely at 34.4 and at 35 weeks, and all the others (18 [72%]) delivered at more than 36 weeks. Conclusions Based on our experience, simple trachelectomy and nodes appear to be a safe fertility-preserving surgery in well-selected patients with small-volume cervical cancer. Obstetric outcome appears favorable.