Celso Abdon Lopes de Mello

p53 As a new prognostic factor for lymph node metastasis in penile carcinoma: Analysis of 82 patients treated with amputation and bilateral lymphadenectomy

PURPOSE Gold standard treatment for invasive penile carcinoma remains amputation and lymphadenectomy. This procedure has high morbidity and new prognostic factors on the incidence of metastasis would help select candidates to lymphadenectomy. Mutations in the p53 gene common in several neoplasms can be related to the prognosis. We studied 82 patients with penile carcinoma staged according to the 1978 TNM system who underwent amputation and bilateral lymphadenectomy to evaluate the prognostic value of immunohistochemical p53 staining in the primary tumor. MATERIALS AND METHODS Immunoreactivity of p53 was studied with other clinicopathological variables, including patient age, stage, histological grade, tumor thickness, lymphatic and venous embolization, corpora cavernosa, corpus spongiosum and urethral infiltration, and human papillomavirus (HPV) status. We also determined its association with lymph node metastasis, the survival rate and the risk of death. In addition, we studied the association of p53 and HPV DNA with prognosis. All slides were reviewed by 1 pathologist. HPV DNA was detected by polymerase chain reaction using GP5/6+ generic primers. p53 expression was measured by immunohistochemical testing with monoclonal Clone DO-7 mouse anti-human p53 protein antibody (Dako A/S, Glostrup, Denmark). The Cox regression hazards method was used for multifactorial analysis. RESULTS Nuclear accumulation of p53 was detected in 34 of 82 samples (41.5%). Clinical lymph node N stage (p = 0.045), lymphatic (p <0.001) and venous (p = 0.04) embolization by neoplastic cells, p53 positivity (p = 0.012) and p53 grade (p = 0.004) were significantly associated with lymph node metastasis. Followup was 0.1 to 453 months (mean 88.7). Multivariate analysis revealed that only lymphatic embolization (relative risk 9.4, 95% confidence interval [CI] 2.8 to 31.6) and p53 positivity (relative risk 4.8, 95% CI 1.6 to 14.9) were independent factors for lymph node metastasis. Patients with negative p53 had significantly better 5 and 10-year overall survival than those in whom tumors stained positive for p53 (64.5% and 54.6% versus 30.2% and 26.4%, respectively, p = 0.009). When tumors were p53 positive and HPV DNA positive, overall survival was worse. Multivariate analysis revealed that only age (relative risk 2.9, 95% CI 1.6 to 5.1) and lymph node metastasis (relative risk 3.2, 95% CI 1.8 to 5.8) were independent risk factors for death. CONCLUSIONS Immunoreactivity of p53 is an independent factor for lymph node metastasis. The association of positive p53 with positive HPV DNA was related to a worse prognosis.

MRP1 expression in CTCs confers resistance to irinotecan-based chemotherapy in metastatic colorectal cancer

Circulating tumor cells are important markers of tumor progression and can reflect tumor behavior in metastatic colorectal cancer (mCRC). Identification of proteins that confer resistance to treatment is an important step to predict response and better selection of treatment for patients. Multidrug resistance‐associated protein 1 (MRP1) and Multidrug resistance‐associated protein 4 (MRP4) play a role in irinotecan‐resistance, and Excision Repair Cross‐Complementation group 1 (ERCC1) expression can confer resistance to platinum compounds. Here, we included 34 patients with mCRC and most of them received FOLFIRI or FOLFOX chemotherapy (91.1%). CTCs were isolated by ISET® Technology and identified in 30 patients (88.2%), with a median of 2.0 CTCs/mL (0–31.0). We analyzed the immunocytochemical expression of MRP1, MRP4 and ERCC1 only in patients who had previously detectable CTCs, accordingly to treatment received (n = 19, 15 and 13 patients, respectively). Among patients treated with irinotecan‐based chemotherapy, 4 out of 19 cases with MRP1 positive CTCs showed a worse progression free survival (PFS) in comparison to those with MRP1 negative CTCs (2.1 months vs. 9.1 months; p = 0.003). None of the other proteins studied in CTCs had significant association with PFS. We analyzed also histological sections of primary tumors and metastases by immunohistochemistry, and found no association with clinicopathological characteristics or with PFS. Our results show MRP1 as a potential biomarker of resistance to treatment with irinotecan when found in CTCs from mCRC patients. This is a small proof‐of‐principle study and these early findings need to be validated in a larger cohort of patients.

Early detection of poor outcome in patients with metastatic colorectal cancer: tumor kinetics evaluated by circulating tumor cells

Background Colorectal cancer (CRC) is the third most prevalent cancer worldwide. New prognostic markers are needed to identify patients with poorer prognosis, and circulating tumor cells (CTCs) seem to be promising to accomplish this. Patients and methods A prospective study was conducted by blood collection from patients with metastatic CRC (mCRC), three times, every 2 months in conjunction with image examinations for evaluation of therapeutic response. CTC isolation and counting were performed by Isolation by Size of Epithelial Tumor Cells (ISET). Results A total of 54 patients with mCRC with a mean age of 57.3 years (31–82 years) were included. Among all patients, 60% (n=32) were carriers of wild-type KRAS (WT KRAS) tumors and 90% of them (n=29) were exposed to monoclonal antibodies along with systemic treatment. Evaluating CTC kinetics, when we compared the baseline (pretreatment) CTC level (CTC1) with the level at first follow-up (CTC2), we observed that CTC1-positive patients (CTCs above the median), who became negative (CTCs below the median) had a favorable evolution (n=14), with a median progression-free survival (PFS) of 14.7 months. This was higher than that for patients with an unfavorable evolution (CTC1− that became CTC2+; n=13, 6.9 months; P=0.06). Patients with WT KRAS with favorable kinetics had higher PFS (14.7 months) in comparison to those with WT KRAS with unfavorable kinetics (9.4 months; P=0.02). Moreover, patients whose imaging studies showed radiological progression had an increased quantification of CTCs at CTC2 compared to those without progression (P=0.04). Conclusion This study made possible the presentation of ISET as a feasible tool for evaluating CTC kinetics in patients with mCRC, which can be promising in their clinical evaluation.

BRAZILIAN CONSENSUS FOR MULTIMODAL TREATMENT OF COLORECTAL LIVER METASTASES. MODULE 3: CONTROVERSIES AND UNRESECTABLE METASTASES

ABSTRACT In the last module of this consensus, controversial topics were discussed. Management of the disease after progression during first line chemotherapy was the first discussion. Next, the benefits of liver resection in the presence of extra-hepatic disease were debated, as soon as, the best sequence of treatment. Conversion chemotherapy in the presence of unresectable liver disease was also discussed in this module. Lastly, the approach to the unresectable disease was also discussed, focusing in the best chemotherapy regimens and hole of chemo-embolization.

EGFR expression in circulating tumor cells from high-grade metastatic soft tissue sarcomas

ABSTRACT Introduction: Soft tissue Sarcomas (STS) are rare malignances, with high mortality rates. Half of patients develop metastasis. The presence of isolated Circulating Tumor Cells (CTCs) and Circulating Tumor Microemboli (CTM) in the blood may be early markers of tumor invasion. Epidermal Growth Factor (EGF) family receptors can also influence this process. Objectives: to quantify CTCs and identify CTM as well as the EGF Receptor (EGFR) protein expression in these cells and correlate with clinical outcome in metastatic STS. Materials and methods: Approximately 8mL of blood was prospectively collected from patients with different types of high-grade STS, before the beginning of chemotherapy. The samples were processed and filtered by ISET (Rarecells, France) for the isolation and quantification of CTCs and CTMs. EGFR expression was analyzed by immunocytochemistry (ICC) on CTCs/ CTMs. Results: We analyzed 18 patients with median age of 49 years (18-77 y). The positivity for EGFR protein expression in CTCs was observed in 93.75% of the patients. This result shows that targeting EGFR positive CTCs from STS origen can be translated in clinical benefit for some patients. In addition, if target therapy is chosen, the EGFR expression in CTCs can be used in follow-up to measure treatment effectiveness. Conclusions: This is the first study to demonstrate the expression of EGFR protein in CTCs from sarcoma patients. It may open an area for future investigations. The next step is to characterize CTCs in a larger cohort of patients to better understand the role of EGFR in sustaining tumor metastasis in sarcomas.

Is early response by 18 F-2-fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography a predictor of long-term outcome in patients with metastatic colorectal cancer?

BACKGROUND Identify in advance responder patients to chemotherapy in metastatic colorectal cancer (CRC) would allow prompt interruption of ineffective therapies in non-responder patients. Hence, predictive markers are sought in numerous trials to detect responder patients, including tumor shrinkage measured by imaging methods. Usually, Response Evaluation Criteria in Solid Tumors (RECIST) is used to evaluate tumor response in metastatic CRC, but these criteria are questionable with use of biological agents associated to chemotherapy. Our aim was correlate early metabolic response by (18)F-2-fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography ((18)FDG-PET-CT) with long-term outcome in metastatic CRC in first-line therapy. METHODS We prospectively evaluated 36 patients with metastatic CRC in first-line treatment with 5-fluorouracil, leucovorin (folinic acid), oxaliplatin (FOLFOX) or 5-fluorouracil, leucovorin (folinic acid), irinotecan (FOLFIRI) associated with cetuximab or bevacizumab. (18)FDG-PET-CT was performed at baseline and after two cycles of chemotherapy. The early metabolic response [standardized uptake value (SUV)] was measured to identify responder and non-responder patients and correlated with overall survival (OS) and progression-free survival (PFS). RESULTS Median age was 58.5 years (range, 41-74 years). PFS was 15.5 months for responder and 13.3 months for non-responder (P=0.42), OS was 55.7 months for responder and not reached for non-responder. There was no correlation between delta-SUV and clinical and pathological variables analyzed. In the subgroup of patients who did not undergo resection of metastasis (45%), PFS was higher for responders (15.3×6.8 months, P=0.02). CONCLUSIONS According to our findings, early response by (18)FDG-PET-CT was not a predictor of long-term outcome for patients with metastatic CRC treated in the first-line chemotherapy with a monoclonal antibody.

Successful treatment of an intra-abdominal desmoid tumor with irinotecan, fluorouracil, and leucovorin plus bevacizumab in a patient with familial adenomatous polyposis

Desmoid tumors, also known as aggressive fibromatosis, are an uncommon neoplasm of soft tissues that do not metastasize but can lead to significant morbidity due a locally aggressive behavior [1, 2]. Although desmoids account for less than 0.1% of all tumors, they are a frequent complication in patients with a hereditary cancer predisposition syndrome, familial adenomatous polyposis (FAP) [3, 4]. FAP is an autosomally dominant disorder caused by mutations of the adenomatous polyposis coli (APC) gene on chromosome 5q21q22. This mutation leads to earlyonset polyposis, characterized by hundreds to thousands of colonic polyps. During life, FAP patients acquire inactivation of the remaining APC gene copy, which accelerates the progression to colorectal cancer [5]. We report a 37-year-old, female patient with metastatic colorectal cancer (mCRC) associated with FAP who also developed a mesenteric desmoid tumor. She was treated with combination of irinotecan, fluorouracil, and leucovorin (IFL) regimen plus bevacizumab as treatment for mCRC and surprisingly had a very good partial response in desmoid tumor. The patient was diagnosed with FAP in 1998 but refused any prophylactic surgery. In February 2008, she was submitted to a subtotal colectomy due to bowel obstruction and a T3N0 right colon adenocarcinoma was diagnosed. No adjuvant treatment was employed. Five months later, she was diagnosed with liver metastasis and treated with a combination of fluorouracil, leucovorin, and oxaliplatin. After 6 months of chemotherapy, CT scan of abdomen showed a partial response in liver lesions and a remarkable new large mass in mesentery which measured 14.0×13.3×10.4 cm (volume 1,936 cm; Fig. 1). At this time we decided to discontinue chemotherapy and perform a CT-guided biopsy of the mesenteric tumor to rule out a second neoplasm. The biopsy was compatible with aggressive fibromatosis. Nuclear beta-catenin immunoreactivity supported the diagnosis of desmoid tumor. She was not considered for surgical procedure due to the morbidity of a wide resection. When she returned for restaging evaluation, CT scan of abdomen showed an increase of liver metastasis size and we decided to treat her with a combination of IFL plus bevacizumab (5 mg/kg) biweekly as second line for mCRC. She tolerated chemotherapy very well. After 3 months of treatment, restaging scan demonstrated a partial response in liver metastasis and a notable decrease in desmoid tumor volume, which measured 12.2×12.1×8.0 cm (volume 1,180 cm). The patient continued with the same combination, and a scan performed after 6 months of therapy L. A. Costa (*) : L. Cezana : T. B. Oliveira :A. L. Dettino : M. F. Fanelli : C. A. Mello Medical Oncology Department, Hospital AC Camargo, São Paulo, Brazil e-mail: leoatem@uol.com.br

Mutation Detection in Tumor-Derived Cell Free DNA Anticipates Progression in a Patient With Metastatic Colorectal Cancer

Background: The observation of tumor-derived cell-free DNA (ctDNA) in plasma brought new expectations to monitor treatment response in cancer patients. Case presentation: In an exploratory case of a 57-year-old man diagnosed with metastatic sigmoid adenocarcinoma, we used a hotspot panel of cancer-associated gene mutations to identify tumor-specific mutations in the primary tumor and metastasis. Results: Five mutations were detected (KRAS, p.Gly12Val; TP53, p.Arg175His; RB1, p.Ile680Thr; ALK, p.Gly1184Glu; and ERBB2, p.Lys860Lys), of which three were detected in both tissue types (primary tumor and metastasis). All five mutations were monitored in the ctDNA of six serial plasma samples. Only KRAS and TP53 mutations were detected at a high frequency in the first plasma sample. After 1 month of chemotherapy the allele frequencies of both mutations fell below the detection limit. From the third month of systemic treatment onward, the allele frequencies of both mutations were detectable in plasma, displaying a continual increase thereafter. The remaining three mutations were not detected in plasma samples. Signs of disease progression in ctDNA during the treatment period were evident while computed tomography (CT) measurements suggested stable metastatic lesions throughout the treatment. Conclusions: Liquid biopsies revealed tumor heterogeneity and predicted tumor progression, demonstrating the potential of ctDNA analysis to be a sensitive and specific tool for monitoring treatment responsivity and for early identification of treatment resistance.

Prognostic implications of phosphatidylinositol 3-kinase/AKT signaling pathway activation in gastric carcinomas

Introduction Activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway plays a critical role in carcinogenesis and resistance to anticancer drugs. In this study, gastric carcinomas (GC) were investigated and statistical analyses were performed concerning the correlation between the clinicopathological features and activation of the PI3K/AKT pathway. Material and methods Immunohistochemistry for p-AKT, p-mTOR and PTEN was performed in 239 GC and 200 non-neoplastic gastric tissues. The clinicopathological parameters were recorded from the medical charts. Statistical significance was defined by a p-value < 0.05. Results High p-AKT expression was observed in 10% of the normal gastric tissue and in 90% of GC, and it was significantly associated with tumor size (p < 0.001), T3/T4 tumors (p < 0.001), and presence of metastases (p = 0.02). Similarly, p-mTOR positivity was found in GC cells, but not in the normal gastric mucosa, and was correlated with perineural invasion (p = 0.02) and T3/T4 tumors (p = 0.03). On the other hand, PTEN expression was weak and focal in the tumor cells, while in the normal gastric tissue this staining was strong and diffuse. Importantly, the expression of p-mTOR and PTEN was associated with overall survival. Conclusions The results of the present study, characterized by the loss of PTEN expression and higher expression of p-AKT and p-mTOR in the majority of tumor cells, apparently are implicated in the carcinogenesis and progression of GC. The identification of p-mTOR and PTEN expression as prognostic factors corroborates the identification and use of potential target drugs that could be more efficient for the treatment of these patients.

Value of mismatch repair deficiency for predicting complete pathologic response to preoperative chemoradiation in rectal carcinoma.

639 Background: The objective of this study is to investigate the value of mismatch repair deficiency (MMR) in rectal cancers as a predictive fator of response to chemoradiation in rectal cancer. Methods: We evaluated a consecutive cohort of 109 patients with locally advanced rectal carcinomas, treated with a total dose of 5,040 cGy of preoperative radiation therapy, concomitant with infusional 5-FU-based chemotherapy. Radical surgery (TME) was performed in all patients. MMR status was determined by imunohistochemistry in tumor tissue from biopsies prior to chemoradiation. The primary endpoint was complete pathologic response (pCR). Results: 63 patients (57.8%) were male, and the mean age was 60 years (range from 28 to 93). The rate of pCR was 18.3%. Among clinical variables (age, sex, distance to dentate line, per-treatment CEA level, and clinical assessment of response), only clinical complete response (cCR) was significant associated with cPR (p=0.009). Only two patients (1.8%) presented tumors with MM...