Daniel Guimarães Tiezzi

Enrichment of regulatory T cells in invasive breast tumor correlates with the upregulation of IL‐17A expression and invasiveness of the tumor

Breast cancer is a leading cause of neoplasia‐associated death in women worldwide. Regulatory T (Treg) and Th17 cells are enriched within some tumors, but the role these cells play in invasive ductal carcinoma (IDC) of the breast is unknown. We show that CD25+CD4+ T cells from PBMCs and tumor express high levels of Foxp3, GITR, CTLA‐4, and CD103, indicating that tumor‐infiltrating Treg cells are functional and possibly recruited by CCL22. Additionally, we observed upregulation of Th17‐related molecules (IL‐17A, RORC, and CCR6) and IL‐17A produced by tumor‐infiltrating CD4+ and CD8+ T lymphocytes. The angiogenic factors CXCL8, MMP‐2, MMP‐9, and vascular endothelial growth factor detected within the tumor are possibly induced by IL‐17 and indicative of poor disease prognosis. Treg and Th17 cells were synchronically increased in IDC patients, with positive correlation between Foxp3, IL‐17A, and RORC expression, and associated with tumor aggressiveness. Therefore, Treg and Th17 cells can affect disease progression by Treg‐cell‐mediated suppression of the effector T‐cell response, as indicated by a decrease in the proliferation of T cells isolated from PBMCs of IDC patients and induction of angiogenic factors by IL‐17‐producing Th17. The understanding of regulation of the Treg/Th17 axis may result in novel perspectives for the control of invasive tumors.

HER-2, p53, p21 and hormonal receptors proteins expression as predictive factors of response and prognosis in locally advanced breast cancer treated with neoadjuvant docetaxel plus epirubicin combination

BackgroundNeoadjuvant chemotherapy has been considered the standard care in locally advanced breast cancer. However, about 20% of the patients do not benefit from this clinical treatment and, predictive factors of response were not defined yet. This study was designed to evaluate the importance of biological markers to predict response and prognosis in stage II and III breast cancer patients treated with taxane and anthracycline combination as neoadjuvant setting.MethodsSixty patients received preoperative docetaxel (75 mg/m2) in combination with epirubicin (50 mg/m2) in i.v. infusion in D1 every 3 weeks after incisional biopsy. They received adjuvant chemotherapy with CMF or FEC, attaining axillary status following definitive breast surgery. Clinical and pathologic response rates were measured after preoperative therapy. We evaluated the response rate to neoadjuvant chemotherapy and the prognostic significance of clinicopathological and immunohistochemical parameters (ER, PR, p51, p21 and HER-2 protein expression). The median patient age was 50.5 years with a median follow up time 48 months after the time of diagnosis.ResultsPreoperative treatment achieved clinical response in 76.6% of patients and complete pathologic response in 5%. The clinical, pathological and immunohistochemical parameters were not able to predict response to therapy and, only HER2 protein overexpression was associated with a decrease in disease free and overall survival (P = 0.0007 and P = 0.003) as shown by multivariate analysis.ConclusionImmunohistochemical phenotypes were not able to predict response to neoadjuvant chemotherapy. Clinical response is inversely correlated with a risk of death in patients submitted to neoadjuvant chemotherapy and HER2 overexpression is the major prognostic factor in stage II and III breast cancer patients treated with a neoadjuvant docetaxel and epirubicin combination.

IL17 Promotes Mammary Tumor Progression by Changing the Behavior of Tumor Cells and Eliciting Tumorigenic Neutrophils Recruitment.

The aggressiveness of invasive ductal carcinoma (IDC) of the breast is associated with increased IL17 levels. Studying the role of IL17 in invasive breast tumor pathogenesis, we found that metastatic primary tumor-infiltrating T lymphocytes produced elevated levels of IL17, whereas IL17 neutralization inhibited tumor growth and prevented the migration of neutrophils and tumor cells to secondary disease sites. Tumorigenic neutrophils promote disease progression, producing CXCL1, MMP9, VEGF, and TNFα, and their depletion suppressed tumor growth. IL17A also induced IL6 and CCL20 production in metastatic tumor cells, favoring the recruitment and differentiation of Th17. In addition, IL17A changed the gene-expression profile and the behavior of nonmetastatic tumor cells, causing tumor growth in vivo, confirming the protumor role of IL17. Furthermore, high IL17 expression was associated with lower disease-free survival and worse prognosis in IDC patients. Thus, IL17 blockade represents an attractive approach for the control of invasive breast tumors.

Co-loaded paclitaxel/rapamycin liposomes: Development, characterization and in vitro and in vivo evaluation for breast cancer therapy

Paclitaxel and rapamycin have been reported to act synergistically to treat breast cancer. Albeit paclitaxel is available for breast cancer treatment, the most commonly used formulation in the clinic presents side effects, limiting its use. Furthermore, both drugs present pharmacokinetics drawbacks limiting their in vivo efficacy and clinic combination. As an alternative, drug delivery systems, particularly liposomes, emerge as an option for drug combination, able to simultaneously deliver co-loaded drugs with improved therapeutic index. Therefore, the purpose of this study is to develop and characterize a co-loaded paclitaxel and rapamycin liposome and evaluate it for breast cancer efficacy both in vitro and in vivo. Results showed that a SPC/Chol/DSPE-PEG (2000) liposome was able to co-encapsulate paclitaxel and rapamycin with suitable encapsulation efficiency values, nanometric particle size, low polydispersity and neutral zeta potential. Taken together, FTIR and thermal analysis evidenced drug conversion to the more bioavailable molecular and amorphous forms, respectively, for paclitaxel and rapamycin. The pegylated liposome exhibited excellent colloidal stability and was able to retain drugs encapsulated, which were released in a slow and sustained fashion. Liposomes were more cytotoxic to 4T1 breast cancer cell line than the free drugs and drugs acted synergistically, particularly when co-loaded. Finally, in vivo therapeutic evaluation carried out in 4T1-tumor-bearing mice confirmed the in vitro results. The co-loaded paclitaxel/rapamycin pegylated liposome better controlled tumor growth compared to the solution. Therefore, we expect that the formulation developed herein might be a contribution for future studies focusing on the clinical combination of paclitaxel and rapamycin.

Breast conserving surgery after neoadjuvant therapy for large primary breast cancer

AIM The aim of this study was to evaluate the safety of breast conserving surgery in patients with breast tumours satisfactorily downstaged after neoadjuvant therapy. METHODS A retrospective cohort study was undertaken to analyze the loco-regional recurrence (LRR) after breast conserving surgery. We enrolled 88 patients with breast cancer subjected to neoadjuvant therapy (NAT group) who achieved an objective response due to neoadjuvant treatment and compared them with 191 patients with early breast cancer (EBC group) who were submitted to primary conserving surgery. Lumpectomy or quadrantectomy with axillary lymph node dissection was performed in all patients who received adjuvant radiotherapy. Systemic adjuvant therapy was offered to all patients. The mean periods of observation were 61.3 months in the NAT group and 67.5 months in the EBC group. RESULTS The mean age was 53 years in the NAT group and 56 years in the EBC group (p=0.04). There was no histological type and histological grade difference between groups. In the NAT group, the mean diameter of residual tumour was lower and the mean volume of breast tissue resection was higher than in the EBC group (p=0.01 and p=0.002, respectively). The ipsilateral recurrence rate was 7.9% in the NAT group and 7.8% in the EBC group (p=0.9). The most important predictive factor of recurrence in the NAT group was the age of patient. CONCLUSION Breast conserving therapy is a safe procedure in satisfactorily downstaged breast cancer after neoadjuvant therapy.

Cirurgia conservadora no câncer de mama

The surgical strategy for breast cancer treatment has changed considerably over the last decade. The breast conserving surgery (BCS) is the standard treatment for early stage breast cancer nowadays. With the current population breast cancer screening programs and the emerging use of systemic neoadjuvant therapy, an increasing number of patients have been eligible to BCS. However, several specific factors must be considered for the therapeutic planning for these patients. This review provides a surgical methodology overview for the BCS in breast carcinoma.

Apoptosis induced by neoadjuvant chemotherapy in breast cancer

Aim: To evaluate the relationship between apoptosis induced by chemotherapy and clinical response in breast cancer. Methods: Apoptosis index (AI), mutant p53 and Bcl‐2 protein expression were evaluated in 44 breast tumour samples from patients submitted to neoadjuvant chemotherapy. Objective response (OR) to primary chemotherapy was observed in 37 patients (84%) and no response (NR) in seven. AI was measured by the rate of apoptotic cells identified using morphological criteria. p53 and Bcl‐2 protein expression were evaluated using an immunoperoxidase staining technique. Results: The median AI change observed between pre‐chemotherapy AI and post‐chemotherapy AI was 0.84 in the OR group and 0.01 in the NR group, (rho = 0.4; p = 0.006). There was no change in Bcl‐2 protein expression following chemotherapy. In the OR group, p53 protein expression was positive in 41.6% of patients before and in 22.2% after chemotherapy (difference = 16.6%; p = 0.03). No change was detected in the NR group. Conclusion: A positive correlation was found between the increase in AI and clinical response to neoadjuvant chemotherapy in locally advanced breast cancer.

Dose-Response Association of CD8+ Tumor-Infiltrating Lymphocytes and Survival Time in High-Grade Serous Ovarian Cancer.

Importance Cytotoxic CD8+ tumor-infiltrating lymphocytes (TILs) participate in immune control of epithelial ovarian cancer; however, little is known about prognostic patterns of CD8+ TILs by histotype and in relation to other clinical factors. Objective To define the prognostic role of CD8+ TILs in epithelial ovarian cancer. Design, Setting, and Participants This was a multicenter observational, prospective survival cohort study of the Ovarian Tumor Tissue Analysis Consortium. More than 5500 patients, including 3196 with high-grade serous ovarian carcinomas (HGSOCs), were followed prospectively for over 24 650 person-years. Exposures Following immunohistochemical analysis, CD8+ TILs were identified within the epithelial components of tumor islets. Patients were grouped based on the estimated number of CD8+ TILs per high-powered field: negative (none), low (1-2), moderate (3-19), and high (≥20). CD8+ TILs in a subset of patients were also assessed in a quantitative, uncategorized manner, and the functional form of associations with survival was assessed using penalized B-splines. Main Outcomes and Measures Overall survival time. Results The final sample included 5577 women; mean age at diagnosis was 58.4 years (median, 58.2 years). Among the 5 major invasive histotypes, HGSOCs showed the most infiltration. CD8+ TILs in HGSOCs were significantly associated with longer overall survival; median survival was 2.8 years for patients with no CD8+ TILs and 3.0 years, 3.8 years, and 5.1 years for patients with low, moderate, or high levels of CD8+ TILs, respectively (P value for trend = 4.2 × 10−16). A survival benefit was also observed among women with endometrioid and mucinous carcinomas, but not for those with the other histotypes. Among HGSOCs, CD8+ TILs were favorable regardless of extent of residual disease following cytoreduction, known standard treatment, and germline BRCA1 pathogenic mutation, but were not prognostic for BRCA2 mutation carriers. Evaluation of uncategorized CD8+ TIL counts showed a near-log-linear functional form. Conclusions and Relevance This study demonstrates the histotype-specific nature of immune infiltration and provides definitive evidence for a dose-response relationship between CD8+ TILs and HGSOC survival. That the extent of infiltration is prognostic, not merely its presence or absence, suggests that understanding factors that drive infiltration will be the key to unraveling outcome heterogeneity in this cancer.

Expression of Hypoxia-inducible factor 1-α and Vascular endothelial growth factor-C in locally advanced breast cancer patients

BACKGROUND: Locally advanced breast cancers are more prevalent in underdeveloped countries. Targeted therapy has been improved to identify hallmarks that are specific to these subtypes of tumors. OBJECTIVES: We aimed to prospectively assess the expression of Hypoxia inducible factor-1 α and vascular endothelial growth factor-C in locally advanced breast cancer patients. METHODS: Thirty women underwent incisional biopsies for the histopathological diagnosis of breast carcinoma and participated in neoadjuvant chemotherapy. The association of Hypoxia inducible factor-1 α and vascular endothelial growth factor-C with age, tumor size, histological grade, clinical staging, hormonal and axillary status, clinical and pathological response after neoadjuvant chemotherapy, expression of estrogen and progesterone receptors, and the presence of c-erbB-2 antigen was studied. RESULTS: Hypoxia inducible factor-1 α expression and Vascular endothelial growth factor-C expression were observed in 66.7% and 63.3% of all patients, respectively, and were marginally associated with each other (p = 0.06). Among the studied variables, only positive axillary status was associated with the presence of HIF-1α (p = 0.02). Complete pathological response was significantly associated (p = 0.04) with the expression of vascular endothelial growth factor-C prior to neoadjuvant chemotherapy. CONCLUSION: We concluded that Hypoxia inducible factor-1 α was associated with a poor prognosis and that vascular endothelial growth factor-C could be used as a predictive factor in locally advanced breast cancer patients with complete pathological response after neoadjuvant chemotherapy.

Câncer de mama: um futuro desafio para o sistema de saúde nos países em desenvolvimento

Na maior parte do mundo, o câncer de mama e a neoplasia maligna que mais acomete a mulher. No Brasil, o cenario nao e diferente. Estima-se que, em 2010, ocorrerao 49.240 novos casos da doenca em nosso pais. De acordo com os dados do GLOBOCAN (International Agency for Research on Cancer), em 2008 foram registrados em todo o mundo 691 mil novos casos de câncer de mama em paises de renda baixa/moderada e 692 mil em paises desenvolvidos. No entanto, o numero total de mortes pela doenca em paises em desenvolvimento foi de 268 mil, em contraste com 189 mil mortes nos paises desenvolvidos. Esses dados resultam em uma taxa de mortalidade/incidencia de 0,38 e 0,27 em paises de baixa/ moderada renda e de alta renda, respectivamente. No relatorio de 2002, foi registrada uma incidencia global de 514 mil casos novos com 221 mil mortes em paises em desenvolvimento (taxa de mortalidade/incidencia de 0,43) e 636 mil casos novos com 190 mil mortes pela doenca em paises desenvolvidos (taxa de mortalidade/incidencia de 0,30). Esses dados permitem estimar o aumento na incidencia de câncer de mama nesses dois grupos de paises. Para os paises de renda baixa/moderada, o aumento da incidencia bruta foi de 25,6% e, nos paises desenvolvidos, de 8,1%. Embora a incidencia de câncer de mama venha sofrendo um leve declinio em alguns paises desenvolvidos em razao da saturacao do sistema de rastreamento da doenca e da reducao do uso indiscriminado da terapia de reposicao hormonal, vem aumentando gradualmente na maioria dos paises em todo o mundo, especialmente em paises em desenvolvimento. Este fato tem sido atribuido principalmente a mudanca no estilo de vida que vem sendo adotado nessas regioes. Estudos com populacoes de imigrantes que saem de regioes de baixa incidencia de câncer de mama para paises com alta taxa de incidencia (como e o caso de imigrantes japoneses ou chineses para os EUA) mostram que os casos de câncer de mama aumentam progressivamente com as sucessivas geracoes. Segundo estimativas da Organizacao Mundial de Saude (OMS), cerca de 7 a 41% de algumas neoplasias malignas podem ser atribuidas ao sobrepeso e obesidade. Adicionalmente, alguns estudos clinicos randomizados demonstraram que modificacoes na dieta com reducao do peso corporal em mulheres na menopausa levam a uma reducao substancial na incidencia de câncer de mama. Essas observacoes sao fortes indicios de que a mudancas no estilo de vida e na dieta pode modificar a epidemiologia da doenca. E de conhecimento de todos que o Brasil e os outros paises em desenvolvimento do mundo sofrem de um problema cronico decorrente da distribuicao irregular de renda entre a