Jorgelina Coppa

Human Tumor-Derived Heat Shock Protein 96 Mediates In Vitro Activation and In Vivo Expansion of Melanoma- and Colon Carcinoma-Specific T Cells

Heat shock proteins (hsp) 96 play an essential role in protein metabolism and exert stimulatory activities on innate and adaptive immunity. Vaccination with tumor-derived hsp96 induces CD8+ T cell-mediated tumor regressions in different animal models. In this study, we show that hsp96 purified from human melanoma or colon carcinoma activate tumor- and Ag-specific T cells in vitro and expand them in vivo. HLA-A*0201-restricted CD8+ T cells recognizing Ags expressed in human melanoma (melanoma Ag recognized by T cell-1 (MART-1)/melanoma Ag A (Melan-A)) or colon carcinoma (carcinoembryonic Ag (CEA)/epithelial cell adhesion molecule (EpCAM)) were triggered to release IFN-γ and to mediate cytotoxic activity by HLA-A*0201-matched APCs pulsed with hsp96 purified from tumor cells expressing the relevant Ag. Such activation occurred in class I HLA-restricted fashion and appeared to be significantly higher than that achieved by direct peptide loading. Immunization with autologous tumor-derived hsp96 induced a significant increase in the recognition of MART-1/Melan-A27–35 in three of five HLA-A*0201 melanoma patients, and of CEA571–579 and EpCAM263–271 in two of five HLA-A*0201 colon carcinoma patients, respectively, as detected by ELISPOT and HLA/tetramer staining. These increments in Ag-specific T cell responses were associated with a favorable disease course after hsp96 vaccination. Altogether, these data provide evidence that hsp96 derived from human tumors can present antigenic peptides to CD8+ T cells and activate them both in vitro and in vivo, thus representing an important tool for vaccination in cancer patients.

Heat shock proteins: biological functions and clinical application as personalized vaccines for human cancer

Heat shock proteins (HSPs) are a large family of proteins with different molecular weights and different intracellular localizations. These proteins undertake crucial functions in maintaining cell homeostasis, and therefore they have been conserved during evolution. Hsp70 and Grp94/gp96, due to their peptide chaperone capacity and their ability to actively interact with professional antigen-presenting cells (APCs), are also endowed with crucial immunological functions. The immunological properties of these proteins and their implications for vaccine in human cancer will be discussed. Immunological and clinical data of phase I/II studies in melanoma and colorectal cancer patients will be reviewed.

Resection versus transplantation for liver metastases from neuroendocrine tumors.

LIVER metastases from neuroendocrine tumors (NET) is the main cause of death for patient with neuroendocrine tumors originating from the intestine and pancreas. In about 90% of patients, the distribution of liver metastases is multifocal and bilateral so that curative liver resection is feasible in no more than 20% of the referred cases. Large liver metastases often cause hormone-related symptoms (carcinoid syndrome) with severe consequences on patient quality of life. Both surgical and medical treatments have been proposed for patients with liver metastases from NET (systemic and intraarterial chemotherapy, somatostatin analogues, interferon therapy) with cumulative patient survival not exceeding 25 to 35% at five years. Resective surgery with curative intent has been associated with an improved 5 year survival in nearly 50% of cases, but the number of eligible patients is low. Total hepatectomy and liver transplantation (OLT) has been advocated for patients with bilateral unresectable symptomatic liver metastases from NET although a clear consensus on stage of disease, pathological subtypes, and patient conditions amenable of transplant candidacy are still lacking. In this report, we describe our experience with 29 patients affected by liver metastases from NET who were treated with either hepatic resections or liver transplantation. Pre-transplantation selection criteria currently applied in our centre are also proposed.

Heat shock proteins and their use as anticancer vaccines

Despite the improvement in the outcome of anticancer therapy achieved during the last few years, several metastatic tumors remain resistant to therapy. This is particularly true for metastatic melanoma, renal and lung carcinoma, and, although to a lesser extent, for colorectal carcinoma. For these

Natural Killer and NK-Like T-Cell Activation in Colorectal Carcinoma Patients Treated with Autologous Tumor-Derived Heat Shock Protein 96

Heat shock proteins (HSPs) are involved in the activation of both adaptive and innate immune systems. Here, we report that vaccination with autologous tumor-derived HSP96 of colorectal cancer patients, radically resected for liver metastases, induced a significant boost of natural killer (NK) activity detected as cytokine secretion and cytotoxicity in the presence of NK-sensitive targets. Increased NK activity was associated with a raise in CD3-CD56+ NK and/or CD3+CD56+ NK-like T cells, displaying enhanced expression of NKG2D and/or NKp46 receptors. Up-regulated expression of CD83 and CD40 and increased interleukin-12 release on stimulation were observed in CD14+ cells from post-HSP96 peripheral blood mononuclear cells, suggesting an indirect pathway of NK stimulation by HSP96-activated monocytes. Additionally, CD3-CD56+ and CD3+CD56+ lymphocytes were found to undergo functional and phenotypic activation on in vitro exposure to HSP96 even in the absence of monocytes, supporting a potential direct activity of HSP96 on these cell subsets. This evidence was confirmed by the specific binding of FITC-conjugated HSP96 to a subset of both CD3-CD56+ and CD3+CD56+ cells in peripheral blood mononuclear cells from colorectal cancer patients. Altogether, these findings identify the activation of the NK compartment as an additional immunologic effect of autologous tumor-derived HSP96 administration in cancer patients.

The challenges of liver transplantation for hepatocellular carcinoma on cirrhosis.

Hepatocellular carcinoma (HCC) is a major cause of cancer mortality worldwide and liver transplantation (LT) has potentials to improve survival for patients with HCC. However, expansion of indications beyond Milan Criteria (MC) and use of bridging/downstaging procedures to convert intermediate‐advanced stages of HCC within MC limits are counterbalanced by graft shortage and increasing use of marginal donors, partially limited by the use of donor‐division protocols applied to the cadaveric and living‐donor settings. Several challenges in technique, indications, pre‐LT treatments and prioritization policies of patients on the waiting list have to be precised through prospective investigations that have to include individualization of prognosis, biological variables and pathology surrogates as stratification criteria. Also, liver resection has to be rejuvenated in the general algorithm of HCC treatment in the light of salvage transplantation strategies, while benefit of LT for HCC should be determined through newly designed composite scores that are able to capture both efficiency and equity endpoints. Innovative treatments such as radioembolization for HCC associated with portal vein thrombosis and molecular targeted compounds are likely to influence future strategies. Accepting this challenge has been part of the history of LT and will endure so also for the future.

The Long-Term Benefit of Liver Transplantation for Hepatic Metastases From Neuroendocrine Tumors

Selection criteria and benefit of liver transplantation for hepatic metastases from neuroendocrine tumors (NETs) remain uncertain. Eighty‐eight consecutive patients with metastatic NETs eligible for liver transplantation according to Milan‐NET criteria were offered transplant (n = 42) versus nontransplant options (n = 46) depending on list dynamics, patient disposition, and age. Tumor burden between groups did not differ. Transplant patients were younger (40.5 vs. 55.5 years; p < 0.001). Long‐term outcomes were compared after matching between groups made on multiple Cox models adjusted for propensity score built on logistic models. Survival benefit was the difference in mean survival between transplant versus nontransplant options. No patients were lost or died without recurrence. Median follow‐up was 122 months. The transplant group showed a significant advantage over nontransplant strategies at 5 and 10 years in survival (97.2% and 88.8% vs. 50.9% and 22.4%, respectively; p < 0.001) and time‐to‐progression (13.1% and 13.1% vs. 83.5% and 89%; p < 0.001). After adjustment for propensity score, survival advantage of the transplant group was significant (hazard ratio = 7.4; 95% confidence interval (CI): 2.4–23.0; p = 0.001). Adjusted transplant‐related survival benefit was 6.82 months (95% CI: 1.10–12.54; p = 0.019) and 38.43 months (95% CI: 21.41–55.45; p < 0.001) at 5 and 10 years, respectively. Liver transplantation for metastatic NETs under restrictive criteria provides excellent long‐term outcome. Transplant‐related survival benefit increases over time and maximizes after 10 years.

Prognostic Role of Pancreatic Metastases From Renal Cell Carcinoma: Results From an Italian Center

BACKGROUND Pancreatic metastasis accounts for 2% to 11% of all mRCC cases. The prognostic value of pancreatic metastases in the era of TTs is unclear. We evaluated outcomes in a cohort of mRCC patients with pancreatic metastases (PmRCC) who were treated with TTs. PATIENTS AND METHODS We retrospectively reviewed the records of 354 mRCC patients treated at our institute between January 2005 and June 2012. Differences in terms of OS between this unselected cohort of mRCC patients and a subgroup of patients with PmRCC were investigated. Kaplan-Meier and log-rank test methods were used to evaluate OS. RESULTS In total, 24 PmRCC (7%) patients were identified, and were compared with a cohort of 330 mRCC patients with metastasis at other sites. Pancreatic metastases were synchronous in 3 patients, and they were metachronous in 11 patients. Surgical resection of pancreatic metastases was performed in 2 (8%) patients. At a maximum follow-up of 89 months (median, 51 months), median OS was 39 months in PmRCC patients, vs. 23 months in the mRCC patient group (P = .0004). CONCLUSION Among mRCC patients treated with TTs, the presence of pancreatic metastasis seems to be associated with a longer survival than the presence of metastasis at other sites.

Fibroblastic reticular cell tumor of the spleen: report of a case and review of the entity

Fibroblastic reticulum cells (FBRCs) are stromal support cells located in the parafollicular area and deep cortex of lymph nodes and in the extrafollicular areas of the spleen and tonsils. We report a case of malignant FBRC tumor of the spleen occurring in a 61-year-old woman. Two years after splenectomy, multiple hepatic lesions were found, which were resected. Histologically, the tumor showed similar morphological features in the spleen as in the liver metastases. There was a whorled pattern of oval and spindle cells in a collagenized background admixed with an inflammatory cell infiltrate composed of lymphocytes and plasma cells. The tumor cells were positive for common muscle actin, smooth muscle actin, and focally for CD68. In situ hybridization for Epstein Barr virus was negative. To the best of our knowledge, this is the first report of malignant FBRC tumor arising in the spleen. The differential diagnosis of splenic tumors with inflammatory pseudotumor-like features is discussed.

Assessment of insulin sensitivity based on a fasting blood sample in men with liver cirrhosis before and after liver transplantation.

Background. Insulin resistance is a key factor in the pathogenesis of hepatogenous diabetes and influences the prognosis of chronic liver diseases. In vivo assessment of insulin resistance in humans is expensive; therefore, surrogate indices based on a fasting plasma glucose and insulin concentrations (HOMA-IS, QUICKI) were proposed. This study aimed to test whether these simple indices are reliable measures of insulin sensitivity in patients with liver cirrhosis before and after liver transplantation (LTx). Methods. HOMA-IS and QUICKI were compared with insulin sensitivity as assessed with the gold standard technique (insulin clamp) in 20 patients with liver cirrhosis, in 36 patients after LTx, and in 25 matched healthy subjects (predominantly men). To test whether these indices may be applied also in prospective studies, 10 patients with liver cirrhosis were studied longitudinally before and 2 years after LTx. Results. Both HOMA-IS and QUICKI were associated with insulin sensitivity in patients with liver cirrhosis (r =0.63, P =0.005 and r =0.60, P =0.009) and in LTx patients (r =0.41, P =0.02 and r =0.46, P =0.05). Both were able to detect the improvement of insulin sensitivity after LTx in the patients studied prospectively. Conclusions. HOMA-IS and QUICKI are simple reliable tools to assess insulin sensitivity in clinical and epidemiologic investigations of chronic liver disease before and after LTx.