In Vitro and In Vivo Efficacy of a Stroma-Targeted, Tumor Microenvironment Responsive Oncolytic Adenovirus in Different Preclinical Models of Cancer.

More than one million women are diagnosed annually worldwide with a gynecological cancer. Most gynecological cancers are diagnosed at a late stage, either because a lack of symptoms, such as in ovarian cancer or limited accessibility to primary prevention in low-resource countries, such as in cervical cancer. Here, we extend the studies of AR2011, a stroma-targeted and tumor microenvironment responsive oncolytic adenovirus (OAdV), whose replication is driven by a triple hybrid promoter. We show that AR2011 was able to replicate and lyse in vitro fresh explants obtained from human ovarian cancer, uterine cancer, and cervical cancer. AR2011 was also able to strongly inhibit the in vitro growth of ovarian malignant cells obtained from human ascites fluid. The virus could synergize in vitro with cisplatin even on ascites-derived cells obtained from patients heavily pretreated with neoadjuvant chemotherapy. AR2011(h404), a dual transcriptionally targeted derived virus armed with hCD40L and h41BBL under the regulation of the hTERT promoter, showed a strong efficacy in vivo both on subcutaneous and intraperitoneally established human ovarian cancer in nude mice. Preliminary studies in an immunocompetent murine tumor model showed that AR2011(m404) expressing the murine cytokines was able to induce an abscopal effect. The present studies suggest that AR2011(h404) is a likely candidate as a novel medicine for intraperitoneal disseminated ovarian cancer.

[High mortality for cervical cancer in young women from the Metropolitan Area of Buenos Aires]. / Elevada mortalidad por cáncer cérvico-uterino en mujeres jóvenes del área Metropolitana de Buenos Aires.

The cervical cancer, which is a reliable indicator of social inequality, remains a major public health issue in Argentina. It is generally accepted that its frequency among young women is low, being the most exposed those over 35 years old. Nevertheless, as gynecologic oncologists, we have been accompanying young patients to their death, mostly women with neither access to screening strategies nor timely or suitable treatment. Such a situation motivated the present analysis of our data on frequency, survival, and demography of cervical cancer collected at the referral cancer hospital of Buenos Aires City. Of 748 cases retrospectively assessed (2007-2011), 84.0% (n = 627) resided in the Metropolitan Area of Buenos Aires; 76.9% (n = 576) were admitted at a locoregionally advanced stage. Regarding tumor size, 53.6% (n = 401) had tumors > 4 cm diameter and 24.2% (n = 181) > 6 cm. The lowest rates of disease-free survival and cause-specific survival were observed for tumor sizes > 6 cm and the age subgroup < 35 years old. Both tumor size and age retained their prognostic value after multivariate analysis adjustment. When focusing in patients under 35 years old, 48% (n = 70) died within 5 years following diagnosis and their probability of surviving 5 years more was < 50%. These figures raise a public health alert on young women with cervical cancer living in the Metropolitan Area of Buenos Aires, which concentrates almost one third of the country population.

An analysis of reported independent prognostic factors for survival in squamous cell carcinoma of the vulva: is tumor size significance being underrated?

OBJECTIVE: To assess independent prognostic factors described in the literature. Thus, to identify different risk groups. METHODS: Review of the records with a diagnosis of primary vulvar squamous cell carcinoma (January/1992-January/2012). INCLUSION CRITERIA: depth of stromal invasion (DSI) >1mm, pathological tumor size >2 cm, and pathological tumor-free margin ≥ 8 mm. Patients who underwent neoadjuvant therapy due to locoregionally advanced vulvar cancer were excluded. All the patients underwent radical, both local and regional, surgery. Adjuvant radiation therapy was administered to all patients with positive nodes. Features of lymph nodes, tumor size, age, grade, lymphovascular space invasion (LVSI), DSI, type of radical surgery, pathological margin distance and stage were evaluated by univariate and multivariate analysis. RESULTS: 194 patients were included. Median age: 67 years. Median follow-up: 62 months. 5-year OS and DFS: 65.5% and 58.2%. Positive lymph nodes were found in 91 (46.9%) patients. After a multivariate analysis, the number of positive lymph nodes, extra-nodal growth, pathologic tumor size and DSI proved to be independent prognostic factors. A high risk group for failure to survive (5y-OS 24%) was identified: tumor size ≥ 6-7.9 cm and DSI >4mm or ≥ 8 cm irrespective of DSI; and extra-nodal growth or ≥2 positive lymph nodes irrespective of tumor size and DSI. CONCLUSIONS: A new high-risk group was identified based on different cutoff values for tumor size, extra-nodal growth and number of positive lymph nodes. This could be very important in the tailored treatment of a specific group of patients with bulky primary tumors and a poorer prognosis.

Argentophilic nucleolus organizer region as a proliferation marker in cervical intraepithelial neoplasia grade 1 of the uterine cervix.

AIM: p16INK4a and argentophilic nucleolus organizer region (AgNOR) can be used as markers for progression of cervical intraepithelial neoplasia grade 1 (CIN1) of the uterine cervix. Our objective was to study the predictive value of the AgNOR technique as a progression marker of CIN1 and its correlation with p16INK4A. MATERIAL AND METHODS: One uterine cervix biopsy from each of 75 patients with diagnosis of CIN1 was selected. All of these patients underwent a second biopsy, and these were also used for the study. RESULTS: The second biopsies showed: regression (20 patients), persistent CIN1 (38 patients), progression to CIN2 (10 patients) and progression to CIN3 (seven patients). p16INK4A showed reactivity in 67 of the 75 first CIN1 biopsies: 12 of the 20 cases that cleared the lesions and the 55 cases with persistent or progressive lesions were positive for p16INK4a (specificity: 40%; sensitivity: 100%; positive predictive value [PPV]: 82%; negative predictive value [NPV]: 100%). Samples with AgNOR areas less than 3.0 µ(2) returned in all cases, but patients whose lesions persisted or progressed to CIN2/CIN3, showed AgNOR areas greater than 3.0 µ(2) in 50/55 cases (specificity: 100%; sensitivity: 91%; PPV: 100%; NPV: 80%). CONCLUSIONS: p16INK4a is expressed in a high percentage of returning lesions. AgNOR might be a better marker of proliferation of CIN1 than p16INK4a (PPV = 100%), which means that a value greater than 3.0 µ(2) indicates the persistence or progression of the lesion. As its NPV is 80%, a value of AgNOR area less than 3.0 µ(2) in CIN1 leaves a margin of doubt about the future behavior of the lesion.

A tumor-stroma targeted oncolytic adenovirus replicated in human ovary cancer samples and inhibited growth of disseminated solid tumors in mice.

Targeting the tumor stroma in addition to the malignant cell compartment is of paramount importance to achieve complete tumor regression. In this work, we modified a previously designed tumor stroma-targeted conditionally replicative adenovirus (CRAd) based on the SPARC promoter by introducing a mutated E1A unable to bind pRB and pseudotyped with a chimeric Ad5/3 fiber (Ad F512v1), and assessed its replication/lytic capacity in ovary cancer in vitro and in vivo. AdF512v1 was able to replicate in fresh samples obtained from patients: (i) with primary human ovary cancer; (ii) that underwent neoadjuvant treatment; (iii) with metastatic disease. In addition, we show that four intraperitoneal (i.p.) injections of 5 × 10(10) v.p. eliminated 50% of xenografted human ovary tumors disseminated in nude mice. Moreover, AdF512v1 replication in tumor models was enhanced 15-40-fold when the tumor contained a mix of malignant and SPARC-expressing stromal cells (fibroblasts and endothelial cells). Contrary to the wild-type virus, AdF512v1 was unable to replicate in normal human ovary samples while the wild-type virus can replicate. This study provides evidence on the lytic capacity of this CRAd and highlights the importance of targeting the stromal tissue in addition to the malignant cell compartment to achieve tumor regression.

Tailoring the treatment of locally advanced squamous cell carcinoma of the vulva: neoadjuvant chemotherapy followed by radical surgery: results from a multicenter study.

OBJECTIVE: To determine the feasibility of performing neoadjuvant chemotherapy (NCH) followed by radical surgery in patients with locally advanced squamous cell carcinoma of the vulva. METHODS: Prospective and multicenter trial. Thirty-five patients with a diagnosis of previously untreated locally advanced squamous cell carcinoma of the vulva were given 4 schemes of cisplatin-based NCH and 1 NCH regimen with single bleomycin. Then, they underwent radical surgery of the vulva if clinical response was 50% or more. Age, NCH schemes used, toxicity, response to treatment, type of radical surgery performed, and clinical outcome were evaluated. RESULTS: Thirty-three patients completed the proposed schemes, and 30 were assessed for radical surgery. Finally, 27 patients underwent radical surgery (radical vulvectomy or radical local excision plus bilateral inguinofemoral lymphadenectomy). In 2 cases of persistent rectal involvement, posterior pelvic exenteration was performed. Moreover, 24 of 27 patients remain with no evidence of disease to date. Toxicity was acceptable. Median age was 62 years (range, 54-72 years). Median follow-up was 49 months (range, 4-155 months). CONCLUSIONS: The use of NCH in selected groups may increase surgical feasibility in initially inoperable patients, thus favoring organ preservation and less extensive resections. Adverse reactions were acceptable, and vulvoperineal deleterious effects that may occur after radiotherapy were consequently avoided.

Oncolytic Adenovirus-Loaded Menstrual Blood Stem Cells Overcome the Blockade of Viral Activity Exerted by Ovarian Cancer Ascites.

Patients with ovarian cancer present peritoneal ascites at recurrence as a marker of disseminated disease and dismal prognosis. Oncolytic immunotherapy is an emerging approach for the treatment of disseminated cancer. In the present work, we constructed a novel oncolytic adenovirus, AR2011, to target malignant ovarian tumors. AR2011 exhibited a clear lytic effect in vitro in human ovarian cancer cell lines and malignant cells obtained from ascitic fluids (AFs) of patients with ovarian cancer. AR2011 activity was neutralized by antibodies present in 31 samples of patient-derived AFs. However, this blockade was overridden by preloading menstrual blood stem cells (MenSCs) with AR2011 (MenSC-AR), since AFs exerted no in vitro inhibitory effect on viral lytic activity under these conditions. Moreover, soluble factors present in AFs act as MenSC chemoattractants. MenSC-AR treatment of nude mice carrying established peritoneal carcinomatosis following administration of human ovarian cancer cells was able to inhibit tumor growth at levels similar to those observed with AR2011 alone. This study demonstrates that MenSCs can be used to override the blockade that AFs exert on viral oncolytic effects.

Defining the concept of locally advanced squamous cell carcinoma of the vulva: a new perspective based on standardization of criteria and current evidence.

The phrase "locally advanced carcinoma of the vulva" has often been mentioned in the literature, though not accurately defined, or even leading to the interpretation overlapping. Grounded on cervical cancer experience, we are able to state that designing a tailored primary strategy based on clinically measurable adverse prognostic factors represents the cornerstone of therapy. This fact urged us to rethink about the real usefulness of the concept of locally advanced squamous cell carcinoma of the vulva. We will refer to this concept as a clinical entity emerging from a rigorous workup which is a valuable guiding tool in the context of a thorough debate about the best primary treatment approach to be used. Furthermore, bulky tumors of the vulva have been associated with a worse prognosis on several occasions. Some authors have questioned the fact that tumor size has not been considered in the staging system. Finally, a standardized definition will help us compare the results obtained, which is extremely necessary given the worldwide low prevalence of this disease.

Elevada mortalidad por cáncer cérvico-uterino en mujeres jóvenes del área metropolitana de Buenos Aires / High mortality for cervical cancer in young women from the Metropolitan Area of Buenos Aires

El cáncer cérvico-uterino, fiel indicador de inequidad social, sigue siendo un grave problema de salud pública en la República Argentina. Se suele afirmar que su frecuencia en mujeres jóvenes es baja y que las más expuestas son aquellas mayores de 35 años. Sin embargo, como ginecólogos oncólogos, con frecuencia acompañamos a morir a mujeres jóvenes que no han tenido acceso a tamizaje ni a tratamiento oportuno y adecuado del cáncer invasor. Esto ha motivado el presente análisis de frecuencia y supervivencia del cáncer cérvico-uterino en el contexto demográfico de las mujeres asistidas en el hospital de referencia en cáncer ginecológico de Buenos Aires. De los 748 casos analizados retrospectivamente (2007-2011), el 84.0% (n = 627) residía en el Área Metropolitana de Buenos Aires y el 76.9% (n = 576) fue admitido en estadios loco-regionalmente avanzados. El 53.6% (n = 401) presentó un diámetro tumoral > 4 cm y el 24.2% (n = 181) > 6 cm. Las tasas más bajas de supervivencia se observaron en tumores > 6 cm y en el subgrupo etario < 35 años. Tanto el tamaño tumoral como la edad conservaron su valor pronóstico tras ser ajustados en el análisis multivariado. En el subgrupo < 35 años, el 48% (n = 70) murió durante los 5 años siguientes al diagnóstico y la probabilidad de sobrevivir otros 5 años fue < 50%. Estos resultados representan una alerta sanitaria sobre la situación de mujeres jóvenes con cáncer cérvico-uterino en el Área Metropolitana de Buenos Aires, la cual concentra casi un tercio de la población del país.

he cervical cancer, which is a reliable indicator of social inequality, remains a major public health issue in Argentina. It is generally accepted that its frequency among young women is low, being the most exposed those over 35 years old. Nevertheless, as gynecologic oncologists, we have been accompanying young patients to their death, mostly women with neither access to screening strategies nor timely or suitable treatment. Such a situation motivated the present analysis of our data on frequency, survival, and demography of cervical cancer collected at the referral cancer hospital of Buenos Aires City. Of 748 cases retrospectively assessed (2007-2011), 84.0% (n = 627) resided in the Metropolitan Area of Buenos Aires; 76.9% (n = 576) were admitted at a locoregionally advanced stage. Regarding tumor size, 53.6% (n = 401) had tumors > 4 cm diameter and 24.2% (n = 181) > 6 cm. The lowest rates of disease-free survival and cause-specific survival were observed for tumor sizes > 6 cm and the age subgroup < 35 years old. Both tumor size and age retained their prognostic value after multivariate analysis adjustment. When focusing in patients under 35 years old, 48% (n = 70) died within 5 years following diagnosis and their probability of surviving 5 years more was < 50%. These figures raise a public health alert on young women with cervical cancer living in the Metropolitan Area of Buenos Aires, which concentrates almost one third of the country population.

Human papillomavirus and mutated H-ras oncogene in cervical carcinomas and pathological negative pelvic lymph nodes: a retrospective follow-up.

The metastasis status of pelvic lymph nodes (PLNs) seems to be a predictive factor of survival. It was suggested that the presence of HPV DNA and other biological markers in PLN may indicate a sub clinical early metastasis. The aim was to describe the prevalence and distribution patterns of HPV DNA and H-ras mutations in intra operatively obtained cervical tumors and PLN. Thirty-seven cervical tumors and 61 lymph node biopsies from 37 patients with cervical cancer were selected. HPV typing and location were performed by PCR/dot blot and in situ hybridization (ISH) respectively. PCR/RFLP was used to scan for mutations in H-ras. Hundred percent of the cervical cancers and 85% of the PLN were HPV positive; co-infection with more than one type was 27%. HPV 16 was detected alone or co-infecting with other types in 84% of tumors and 46% of PLN; the second most frequent viral type was HPV 18 (tumor: 27%; PLN: 20%). In PLN, HPV was located in nuclei or/and cytoplasm of lymphocytes, macrophages, endothelial, and /or stromal cells. H-ras mutations were identified in 5/24 (21%) of patients with cervical tumors showing poor or moderated differentiation. HPV DNA in histological tumor-free PLN not necessary indicate metastasis, but it may be associated to an active immune reaction. Mutated H-ras is probably involved in cervical carcinogenesis and its detection in tumor and metastasis free PLN may be related to early metastasis or recurrence in at least a subset of poorly differentiated cervical tumors.

Fallas en la confección del consentimiento informado oncoginecológico / Deficiencies in the confection of an oncogynecologic informed consent

In recent decades, the relationship between patients and physicians has been evolving from the traditional paternalistic relationship in which physicians made choice for patients, to a more equal relationship of shared decision making in which physicians provide information and counseling that allows competente adult patients to make their own choices. The process by which physician and patients make decisions together, is summarized in the concept "informed consent"...In this process, the physician educates the patient and her family and the patient chooses treatment based on reasonable medical alternatives. All these aspects are considered in the article

El correcto seguimiento de la enferma portadora de cáncer de mama : importancia de la motivación / The correct follow-up of the patient with breast cancer : importance of the motivation

Definimos lo que entendemos por correcto seguimiento de la paciente con cáncer de mama, fijamos los objetivos entre los que se destacan la continuidad de la relación médico-paciente, el apoyo psicoterapéutico y la colaboración del servicio social. Ordenamos la metodología y nos abocamos al texto de la motivación y al contenido de la información adaptándola al nivel y capacidad de entendimiento de la enferma. Evaluamos a los responsables del seguimiento, analizamos las dificultades del mismo. Y destacamos las condiciones del médico encargado de la conducción del seguimiento.