SUMMARY We describe the landscape of genomic alterations in cutaneous melanomas through DNA, RNA, and protein-based analysis of 333 primary and/or metastatic melanomas from 331 patients. We establish a framework for genomic classification into one of four subtypes based on the pattern of the most prevalent significantly mutated genes: mutant BRAF, mutant RAS, mutant NF1, and Triple-WT (wild-type). Integrative analysis reveals enrichment of KIT mutations and focal amplifications and complex structural rearrangements as a feature of the Triple-WT subtype. We found no significant outcome correlation with genomic classification, but samples assigned a transcriptomic subclass enriched for immune gene expression associated with lymphocyte infiltrate on pathology review and high LCK protein expression, a T cell marker, were associated with improved patient survival. This clinicopathological and multidimensional analysis suggests that the prognosis of melanoma patients with regional metastases is influenced by tumor stroma immunobiology, offering insights to further personalize therapeutic decision-making.

Cluster of differentiation (CD)8 ؉ T cells exist as naive, central memory, and effector memory subsets, and any of these populations can be genetically engineered into tumor-reactive effector cells for adoptive immunotherapy. However, the optimal subset from which to derive effector CD8 ؉ T cells for patient treatments is controversial and understudied. We investigated human CD8 ؉ T cells and found that naive cells were not only the most abundant subset but also the population most capable of in vitro expansion and T-cell receptor transgene expression. Despite increased expansion, naive-derived cells displayed minimal effector differentiation, a quality associated with greater efficacy after cell infusion. Similarly, the markers of terminal differentiation, killer cell lectin-like receptor G1 and CD57, were expressed at lower levels in cells of naive origin. Finally, naive-derived effector cells expressed higher CD27 and retained longer telomeres, characteristics that suggest greater proliferative potential and that have been linked to greater efficacy in clinical trials. Thus, these data suggest that naive cells resist terminal differentiation, or "exhaustion," maintain high replicative potential, and therefore may be the superior subset for use in adoptive immunotherapy. (Blood. 2011;117(3):808-814) IntroductionIt is now possible to genetically engineer human T lymphocytes to express virtually any known gene, including genes encoding T-cell receptors (TCRs) or chimeric antigen receptors (CARs), to provide the desired T-cell specificity. These gene-engineered lymphocytes hold the promise to treat infectious diseases and cancer, 1-10 but the appropriate substrate cells to use is understudied, and most current protocols simply transfer genes into "bulk" peripheral blood mononuclear cells (PBMCs). Varieties of data indicate that T cells, and in particular CD8 ϩ T cells, are worthy substrates for gene engineering. However, CD8 ϩ T cells in peripheral blood are themselves a complex mixture, composed of at least 3 major subsets-naive (T N ), central memory (T CM ), and effector memory (T EM )-each having different functional qualities.The optimal subset to engineer for adoptive immunotherapy is controversial. 8,[11][12][13][14] Memory CD8 ϩ T-cell subsets are more studied than naive cells, because antigen-specific clones can be found at increased frequencies. In a landmark paper, Berger et al 13 have shown in macaques that effector cells derived from T CM rather than T EM possess greater ability to survive and establish immunologic memory after infusion. This finding is consistent with data comparing memory subsets in mice. 15 However, T N cells were not explicitly analyzed in these reports, 12 an omission that might be important given recent findings in mice that naive T cells convey more antitumor activity than memory cells. 12 Short of conducting a series of clinical trials, the decision of which CD8 ϩ T-cell subset to use for clinical protocols will depend on the cell phenotypes that result from transduction of T...

The mechanisms which allow cancer cells to adapt to the typical tumor microenvironment of low oxygen and glucose and high lactate are not well understood. GPR81 is a lactate receptor recently identified in adipose and muscle cells that has not been investigated in cancer. In the current study, we examined GPR81 expression and function in cancer cells. We found that GPR81 was present in colon, breast, lung, hepatocellular, salivary gland, cervical and pancreatic carcinoma cell lines. Examination of tumors resected from pancreatic cancer patients indicated that 94% (148/158) expressed high levels of GPR81. Functionally, we observed that the reduction of GPR81 levels using shRNA mediated silencing had little effect on pancreatic cancer cells cultured in high glucose, but led to the rapid death of cancer cells cultured in conditions of low glucose supplemented with lactate. We also observed that lactate addition to culture media induced the expression of genes involved in lactate metabolism including monocarboxylase transporters in control, but not in GPR81 silenced cells. In vivo, GPR81 expression levels correlated with the rate of pancreatic cancer tumor growth and metastasis. Cells in which GPR81 was silenced showed a dramatic decrease in growth and metastasis. Implantation of cancer cells in vivo was also observed to lead to greatly elevate levels of GPR81. These data support that GPR81 is important for cancer cell regulation of lactate transport mechanisms. Furthermore, lactate transport is important for the survival of cancer cells in the tumor microenvironment.

This is the first large-scale validation of the AJCC 8th edition staging system for pancreatic cancer. The revised system provides discrimination similar to that of the 7th-edition system. However, the 8th-edition system allows for finer stratification of patients with resected tumors according to extent of nodal involvement.

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