Purpose Lymphopenia as a likely index of poor systemic immunity is an independent predictor of inferior outcome in clear renal cell carcinoma (ccRCC). We sought to evaluate the prognostic relevance of preoperative absolute lymphocyte count (ALC) in a cohort of papillary renal cell carcinoma (PRCC) patients. Materials & Methods A prospectively maintained, renal cancer database was analyzed. Patients with preoperative ALC, within 3 months prior to surgery, were eligible for the study. Those with multifocal or bilateral renal tumors were excluded. Correlations between ALC and age, gender, smoking, Charlson comorbidity index (CCI), pathologic (pT) stage, PRCC subtype, and TNM stage were evaluated. Differences in overall survival (OS) & cancer-specific survival (CSS) by ALC status were assessed using the log–rank test and cumulative incident estimators, respectively. Cox proportional hazards modeling was used for multivariable analyses (MVA). Results 192 patients met the inclusion criteria As a continuous variable, preoperative ALC was associated with higher TNM stage (p=0.001) and older age (p=0.01). As a dichotomous variable, lymphopenia (<1,300 cells/μl) was associated with higher TNM stage (p=0.003). On MVA, controlling for covariates, after a median follow up of 37.3 months, lymphopenia was associated with inferior OS (HR=2.3 [95%CI 1.2–4.3], p=0.011) and trended to significance for CSS (p=0.071). Among non-metastatic, lymphopenic patients, OS at 37.5 months was shorter compared to those with normal ALC (83% vs. 93%, p=0.0006). Conclusions In patients with PRCC, lymphopenia is associated with lower survival independent of TNM stage, age, and histology. ALC may provide an additional pre-operative prognostic factor.

Distinct intracellular pathways are involved in regulated and constitutive protein secretion from neuronal and endocrine cells, yet the peptide signals and molecular mechanisms responsible for targeting and retention of soluble proteins in secretory granules are incompletely understood. By using confocal microscopy and subcellular fractionation, we examined trafficking of the neuronal and endocrine peptide precursor VGF that is stored in large dense core vesicles and undergoes regulated secretion. VGF cofractionated with secretory vesicle membranes but was not detected in detergent-resistant lipid rafts. Deletional analysis using epitope-tagged VGF suggested that the C-terminal 73-amino acid fragment of VGF, containing two predicted ␣-helical loops and four potential prohormone convertase (PC) cleavage sites, was necessary and sufficient with an N-terminal signal peptide-containing domain, for large dense core vesicle sorting and regulated secretion from PC12 and INS-1 cells. Further transfection analysis identified the sorting sequence as a compact C-terminal ␣-helix and embedded 564 RRR 566PC cleavage site; mutation of the 564 RRR 566 PC site in VGF-(1-65): GFP:VGF-(545-617) blocked regulated secretion, whereas disruption of the ␣-helix had no effect. Mutation of the adjacent 567 HFHH 570 motif, a charged region that might enhance PC cleavage in acidic environments, also blocked regulated release. Finally, inhibition of PC cleavage in PC12 cells using the membrane-permeable synthetic peptide chloromethyl ketone (decanoyl-RVKR-CMK) blocked regulated secretion of VGF. Our studies define a critical RRR-containing C-terminal domain that targets VGF into the regulated pathway in neuronal PC12 and endocrine INS-1 cells, providing additional support for the proposed role that PCs and their cleavage sites play in regulated peptide secretion.Regulated secretion of polypeptides from neuronal and endocrine cells, which relies on packaging of proteins into the appropriate vesicle pools within the cell, is a critical control point for regulating peptide levels and ultimately function. Polypeptide motifs that target cell surface proteins to specific vesicle populations or to discrete locations within the cell, such as the basolateral or apical surface of a polarized epithelial cell, have been relatively well characterized (1). Protein domains that are responsible for targeting soluble proteins into the regulated release pathway appear to be more heterogeneous, and thus generalized sorting domains and consensus motifs have been more difficult to identify. Perhaps as a consequence, several models that explain targeting into the regulated secretory pathway by selective sorting and/or selective retention have been proposed (2-5). Following signal sequence-directed translocation into the lumen of the endoplasmic reticulum, segregation of proteins into the constitutive or regulated pathways is suspected to occur in the trans-Golgi network (TGN). 4 Constitutive secretory vesicles transit directly from the TGN to the plasma membran...

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Introduction and Objective Sarcomatoid differentiation in renal cell carcinoma (sRCC) is associated with a very poor prognosis. The identification of genetic alterations that drive this aggressive phenotype could aid in the development of more effective targeted therapies. In this study, we aimed to pinpoint unique copy number alterations (CNAs) in sRCC when compared to classical RCC subtypes. Methods Genomic copy number analysis was performed using single nucleotide polymorphism (SNP)-based microarrays on tissue extracted from the tumors of 81 patients who underwent renal mass excision, including 17 with sRCC. Results sRCC tumors exhibited significantly higher numbers of CNAs when compared to clear cell (ccRCC), papillary (pRCC) and chromophobe RCC (chRCC) (mean 18.0 vs. 5.8, 6.5, and 7.2, respectively; p <0.0001). Copy number losses of chromosome arms 9q, 15q, 18p/q, and 22q and gains of gains of 1q and 8q occurred in a significantly higher proportion of sRCC tumors compared to the other 3 histologies. Patients with sRCC tumors demonstrated significantly worse overall survival when compared to those without sRCC on Kaplan-Meier analysis (p=0.0001). Patients with 9 or more CNAs also demonstrated significantly worse overall survival compared to those with fewer than 9 CNAs (p=0.004). Conclusions Sarcomatoid differentiation in RCC is associated with a high rate of chromosomal imbalances with losses of 9q, 15q, 18p/q and 22q, and gains of 1q and 8q occurring at significantly higher frequencies in comparison to non-sRCC tumors. Identification of candidate driver genes or tumor suppressor loci within these chromosomal regions may help identify targets for future therapies.