Abstract
Purpose of the study: Epithelial ovarian cancer (OC) is one of the most common gynecologic malignancies and the fifth most frequent cause of cancer death in women. Paclitaxel and Carboplatin ...chemotherapy (CT) remains the standard treatment, but tumor relapses in approximately 80% of patients, mostly due to drug resistance. Novel therapeutic options are therefore strongly required. Recently, human Anti-Müllerian Hormone type II Receptor (AMHRII) has been identified as a potential target, but up to date the AMHRII expression in ovarian tumors has been poorly documented. A humanized glyco-engineered monoclonal antibody (mAb) called 3C23K with higher Fc effector function has been developed. The aim of this study is to investigate the potential efficacy of this mAb by assessing AMHRII expression and analyzing the localization and amount of mAb responsive immune effectors in OC patient and in a patient-derived xenograft (PDX) model.
Experimental procedures: AMHRII expression was prospectively studied in fresh biopsies (primary ovarian tumors, peritoneal carcinomatosis and ascites) from a cohort of 28 patients, by means of both flow cytometry (FC) and immunofluorescence (IF). We have characterized the intratumoral immune infiltrate by FC and determined the localization of immune cells within the tumor by IF. Similar experiments were performed on an undifferentiated highly proliferative serous ovarian adenocarcinoma PDX model, in parallel to in vivo efficacy study of 3C23K alone or in combination with standard CT. The antitumor activity of infiltrating immune cells isolated from OC patients (peritoneal washing) was assessed using red blood cells as target and a glyco-engineered anti-D Rhesus mAb. The antibody-dependent cell cytotoxicity (ADCC) and the antibody-dependent cell phagocytosis (ADCP) abilities of 3C23K against AMHRII+ cells (COV434-RII) were analyzed by IF using monocyte-derived macrophages (MDM) from blood of healthy donors as effector cells.
Summary of the data: Here, we show that among the 28 ovarian tumors tested, 19 patients (68%) were positive for AMHRII with at least 15% of tumor cells being positive for AMHRII. OC are largely composed of (CD45+) immune cells mostly composed of tumor associated macrophages (TAMs) and tumor infiltrating lymphocytes (TILs). TAMs highly express M2-like markers, such as CD206, but also strongly express the Fcγ III receptor (CD16). Unlike TILs, which accumulate in the peritumoral stroma, CD16+ TAMs are found in both the stroma and the tumor islets. Moreover, tumors infiltrating immune cells are able to induce a mAb-dependent cytotoxicity ex vivo. In parallel, in vivo experiments have shown an intermediate efficacy of the 3C23K antibody administered alone and a significantly increased antitumor effect in combination with CT including a prolonged survival. IF and FC analyses showed high TAM infiltration compatible with induction of ADCP by 3C23K in this model. Furthermore, 3C23K mAb induces ADCC and/or ADCP of COV434-RII cells by MDM likely through CD16 interaction, either in the absence or presence of saturating concentrations of Ig.
Conclusions: We have confirmed the expression of AMHRII in 2/3 of OC patients. MAb responsive immune effectors and particularly macrophages largely infiltrate OC and are found in contact with malignant cells. Targeting AMHRII by the 3C23K mAb dramatically increases the efficacy of a standard CT in vivo. 3C23K appears to act through macrophage-induced ADCC and/or ADCP of tumor cells and current studies are designed to refine its mechanism of action. Our results indicate that most ovarian tumors display optimal conditions for a 3C23K-based immunotherapy. Hence, the combination of 3C23K with CT could bring additional therapeutic benefit in OC patients compared to standard chemotherapy.
Citation Format: Houcine Bougherara, Fariba Némati, Christophe De Romeuf, Jean-Marc Barret, Gérald Massonnet, Marie-Aude Le Frère-Belda, Jean-François Prost, Didier Decaudin, Emmanuel Donnadieu. Ex vivo evaluation of an anti-Müllerian hormone type II receptor humanized antibody with optimized Fc effector function in ovarian cancer. abstract. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A01.
Abstract only
e15558
Background: Pelvic lymphadenectomy is associated with a significant risk of lower-limb lymphedema. In this proof-of-concept study, we evaluated the feasibility of identifying the ...lower-limb drainage nodes (LLDNs) during pelvic lymphadenectomy for endometrial cancer. Secondary objectives were to map lower-limb drainage and to assess the diagnostic value of our mapping technique. Methods: Prospective study of patients with endometrial cancer requiring pelvic lymphadenectomy, with no neoadjuvant radiotherapy or chemotherapy and no history of lower-limb surgery. A radiopharmaceutical was injected into both feet on the day before surgery. LLDNs were identified using preoperative lymphoscintigraphy and intraoperative isotopic probe detection then removed before complete pelvic lymphadenectomy. LLDNs and pelvic lymphadenectomy specimens underwent separate histological analysis. Results: Of the 12 patients with early-stage endometrial cancer, 10 underwent preoperative lymphoscintigraphy, which consistently identified inguinal, femoral and pelvic LLDNs. The intraoperative detection rate was 83% (10/12). The 2 patients with no LLDNs detected intraoperatively had lymphoscintigraphy-detected LLDNs. Median number of hot nodes per patient was 5 (3-7) on the right and 3 (2-6) on the left. Of 107 LLDNs, 106 were in the external iliac area, including 38 in the lateral group and 45 in the intermediate and medial groups. None of the patients had node metastases at any site. No early complications related to the technique occurred. Conclusions: Our mapping technique appears feasible, safe, and associated with a high LLDN identification rate. LLDN mapping may allow the preservation of LLDNs, thereby decreasing the risk of lower-limb lymphedema and improving quality of life.
Abstract only
5098
Background: Poor data exist on clinico-pathological description of endometrial cancer (EC) in Lynch syndrome (LS) compared with sporadic ones. To evaluate the clinico-pathological ...findings of Lynch-related EC to establish histological criteria to discriminate familial and sporadic EC and to decide the optimal management of patients. Methods: Retrospective study of prospective cohort of patients with LS in our institution from 1999 to 2011. We identified and described all cases of endometrial cancers. Management and follow-up were detailed. Results: Of a cohort of 126 patients with LS, 10 women developed endometrial carcinoma. The median age at diagnosis was 51 years (41-58). Five patients had an identified mutation (2 hMLH1, 2 hMSH2 and 1 hMSH6). In 9 cases, EC was the first Lynch-related tumor to occur. No patient developed ovarian cancer. All, except 2 patients (1 serous carcinoma and 1 clear cell carcinoma), had endometrioid adenocarcinoma (80%). Tumor grade was grade 1 in 3 patients, grade 2 in 5 and grade 3 in 2 patients. Forty per cent of patients had lymphovascular space involvement (LVSI). The FIGO stages were as follows: stage IA (n=7), stage IB (n=2) and stage IIIC (n=1). Four in ten patients had tumor located in the lower uterine segment. With a median follow-up of 14 months (range 9 – 40 months), recurrence occurred in one patient with a stage IB grade 2 endometrioid adenocarcinoma with LVSI. Conclusions: EC in LS is characterized by early age at onset, localization in lower uterine segment, and high rate of LVSI. Other data on histology and survival do not differ from sporadic cancers. These results suggest that we can consider conservative treatment in patients with good prognosis tumors. Further studies are required.
The INK4a/ARF locus encodes 2 cell cycle regulatory proteins: p16 and p14(ARF). P16 inhibits the activities of cdks, which maintain the retinoblastoma protein (pRb) in its active hypophosphorylated ...state. P14(ARF) blocks MDM2-induced p53 degradation and transactivational silencing. In this study, we investigated the expression of p16 and p14(ARF) in reference human urothelium and in 51 urothelial carcinomas (UCs) of all stages and grades, by reverse transcription-polymerase chain reaction (RT-PCR). Patterns of p14(ARF) and p16 expression were compared with each other and then with patterns of p53 and pRb protein expression, respectively, as determined by immunohistochemistry. P14(ARF) and p16 mRNAs were present at low levels or were undetectable in reference urothelia and in most superficial tumors, whereas they were present at high levels in a subset of tumors of advanced stage and high grade. The expression profiles of these 2 mRNAs were correlated in all but 4 cases, indicating that the 2 INK4a products may have nonredundant functions. Forty-six of the 51 tumors (90%) presented changes to or a lack of activation of the p14(ARF)-p53 pathway and were p53 positive (n = 10), p14(ARF) negative (n = 23), or both p53 positive and p14(ARF) negative (n = 13), suggesting that these 2 components of the pathway may be altered or nonactivated. Markedly high levels of p16 mRNA (n = 5) were associated with the absence of pRb expression, with the exception of 1 case in which the p16 gene contained a deletion mutation. A lack of p16 mRNA or low levels of this mRNA were associated with pRb detection in all but 1 case. In invasive UCs, the p16-pRb pathway, the p14(ARF)-p53 pathway, or in many cases both pathways were altered or not activated, demonstrating the involvement of these pathways in invasive bladder tumorigenesis.
It is well known that several metals, such as lead, mercury, cadmium, and vanadium, can mimic the effects of estrogens (metallo-estrogens). Nevertheless, there are only a few studies that have ...assessed the effects of toxic metals on the female genital tract and, in particular, endometrial tissue. In this context, we measured the concentrations of several trace elements in human endometrial tissue samples from individuals with hyperplasia or adenocarcinoma and in normal tissues. Hyperplasic endometrial tissue has a 4-fold higher concentration of mercury than normal tissue. Mercury can affect both the AhR and ROS signaling pathways. Thus, we investigated the possible toxic effects of mercury by in vitro studies. We found that mercury increases oxidative stress (increased HO1 and NQO1 mRNA levels) and alters the cytoskeleton in the human endometrial Ishikawa cell line and to a lesser extent, in the "less-differentiated" human endometrial Hec-1b cells. The results might help to explain a potential link between this metal and the occurrence of endometrial hyperplasia.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Oncogenetic testing is now part of standard management in high grade ovarian cancer, including at least mutational status of BRCA1/BRCA2 genes. If necessary, tumor genetic testing is followed by ...constitutional testing to either confirm the constitutional origin of variants identified in BRCA1/2 genes or detect variants in other predisposition genes. The whole process including prescription of tumoral testing, retrieval of analysis report and communication of results must be formalized, as well as information on possible consequences of the results for the patient and her family. Tumor material must meet criteria of size and cellularity to allow high-quality analysis. These samples are processed during the preanalytical phase with two major steps : time of cold ischemia and fixation. Only pathogenic (Class V) and likely pathogenic (Class IV) variants shown in tumor tissue are mentioned in the report. Currently, only BRCA1 and BRCA2 genes are routinely studied but, in the future, analysis will be extended to other genes involved in homologous recombination repair. In patients without BRCA mutation, other biomarkers reflecting sensitivity to PARP inhibitors, such as HRD scores (homologous recombination deficiency) that appeared recently, will have to be implemented in routine practice in order to better select patients for these treatments and choose optimal therapy.
The sentinel lymph node (SLN) biopsy has been proposed for the cancers of the uterus in order to optimize the diagnosis of lymphatic metastases and micrometastases in early stage tumors. Patients ...with early invasive cervical (n = 8) or endometrial (n = 15) cancers were enrolled. A lymphoscintigraphy was carried out before the intervention. Intraoperative SLN identification was performed with blue dye combined to a handheld gamma probe detection. Non-sentinel pelvic nodes were separately cleared out. SLNs were examined with frozen sections, permanent sections with hematoxylin-eosin staining and further serial sections with immunohistochemistry if negative. Six cervical cancer patients and 13 endometrial cancer patients had a positive lymphoscintigraphy, showing in 5 patients extra-iliac SLN(s). The intraoperative detection was successful in 6 cervical cancer patients and 14 endometrial cancer patients. The higher detection rate was obtained with the isotopic method. Most of the SLNs were ilio-obturator. Four endometrial cancer patients had a lymphatic spread, only involving the SLN in each case. No false negative SLN has been noted. SLN biopsy appears feasible in cervical and endometrial cancers. This procedure could improve the lymphatic evaluation of these cancers.
The
INK4a/ARF locus encodes 2 cell cycle regulatory proteins: p16 and p14
ARF. P16 inhibits the activities of cdks, which maintain the retinoblastoma protein (pRb) in its active hypophosphorylated ...state. P14
ARF blocks MDM2-induced p53 degradation and transactivational silencing. In this study, we investigated the expression of p16 and p14
ARF in reference human urothelium and in 51 urothelial carcinomas (UCs) of all stages and grades, by reverse transcription-polymerase chain reaction (RT-PCR). Patterns of p14
ARF and p16 expression were compared with each other and then with patterns of p53 and pRb protein expression, respectively, as determined by immunohistochemistry. P14
ARF and p16 mRNAs were present at low levels or were undetectable in reference urothelia and in most superficial tumors, whereas they were present at high levels in a subset of tumors of advanced stage and high grade. The expression profiles of these 2 mRNAs were correlated in all but 4 cases, indicating that the 2
INK4a products may have nonredundant functions. Forty-six of the 51 tumors (90%) presented changes to or a lack of activation of the p14
ARF-p53 pathway and were p53 positive (n = 10), p14
ARF negative (n = 23), or both p53 positive and p14
ARF negative (n = 13), suggesting that these 2 components of the pathway may be altered or nonactivated. Markedly high levels of p16 mRNA (n = 5) were associated with the absence of pRb expression, with the exception of 1 case in which the p16 gene contained a deletion mutation. A lack of p16 mRNA or low levels of this mRNA were associated with pRb detection in all but 1 case. In invasive UCs, the p16-pRb pathway, the p14
ARF-p53 pathway, or in many cases both pathways were altered or not activated, demonstrating the involvement of these pathways in invasive bladder tumorigenesis.
PDF
