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Prostate cancer (PCa) is characterized by alterations in lipid metabolism, exemplified by increased rates of de novo lipogenesis and elongation by fatty acid elongase enzymes known as ELOVLs (elongation of very long chain fatty acids). We have previously shown that the ELOVL family members ELOVL2, 5, and 7 are Androgen Receptor (AR)-regulated proteins; and that ELOVL5 is overexpressed in both primary and castration-resistant prostate cancer. To confirm the role of ELOVL5 activity in prostate carcinogenesis, we generated genetically engineered mouse models (GEMM) with deletion of Elovl5, validated by immunohistochemistry, in combination with the overexpression of c-Myc, an oncogene known to affect lipid metabolism in cancer cells. As expected, we observed alterations in the lipid profile of the murine prostate with the partial deletion of Elovl5, as determined by a significant reduction in levels of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) with long-chain fatty acids (22 carbons) and increased levels of PE with short-chain fatty acids (14 carbons), suggesting an overall shortening on acyl chains of the most common phospholipid species. Interestingly, we also observed that the heterozygous deletion of Elovl5 reduced the expression levels of the AR on the anterior lobe of the murine prostate, in comparison to tissue from Elovl5 wild type mice. Preliminary data also indicates a modulation in prostate volume when Elovl5 is genetically deleted in comparison to tissue from wild type mice. Heterozygous Elovl5 deletion did not affect healthy tissue.To explore biological effects, we generated a 3D organoid line derived from the prostate of mice with homozygous deletion of Elovl5 and overexpression of c-Myc. We found that Elovl5 knock-out decreased organoid diameter and proliferation of c-Myc-driven cells compared to Elovl5 WT. This effect was potentiated when organoids were treated with Enzalutamide, suggesting that targeting Elovl5 may sensitize prostate cells to anti-androgen therapy.Our findings suggest lipid elongation via ELOVL5 as a potential therapeutic target for PCa. Citation Format: Silvia D. Rodrigues, Caroline Ribeiro, Isadora Teixeira, Hubert Pakula, Giuseppe N. Fanelli, Fabio Socciarelli, Lisa Butler, Johannes Swinnen, Massimo Loda. ELOVL5 affects prostate cancer cell proliferation and modulates the AR pathway. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6039.

The associated publication reports proteogenomic analysis of non-small cell lung cancer, where we identified molecular subtypes with distinct immune evasion mechanisms and therapeutic targets, and validated our classification method in separate clinical cohorts. This protocol describes histological, tertiary lymphoid structure (TLS), and immunohistochemical evaluation of clinical samples. Specifically, immunohistochemistry was performed for PD-L1, CD3, and CD8 on tumor microarrays (TMAs) derived from formalin-fixed paraffin embedded (FFPE) samples.

The associated publication reports proteogenomic analysis of non-small cell lung cancer (NSCLC), where we identified molecular subtypes with distinct immune evasion mechanisms and therapeutic targets, and validated our classification method in separate clinical cohorts. This protocol describes sections of the bioinformatics analysis of the multi-omics data, namely, data analysis and processing for panel sequencing, identification of cancer- and driver-related proteins in proteomics data, proteogenomics search, and machine learning-based classifiers for NSCLC subtyping. Specifically, a cohort classifier was built using support-vector machine-recursive feature elimination (SVM-RFE) algorithm applied to in-depth proteomics data from a cohort of 141 samples. The classifier was then validated in three external datasets. Another classifier, suitable for single-sample subtyping, was built using k-top scoring pairs (k-TSP) algorithm applied to label-free data from a cohort of 136 samples. The k-TSP-based classifier was validated in two independent cohorts and an additional external dataset.

The associated publication reports proteogenomic analysis of non-small cell lung cancer (NSCLC), where we identified molecular subtypes with distinct immune evasion mechanisms and therapeutic targets, and validated our classification method in separate clinical cohorts. The in-depth proteomic analysis pointed to a potential role of STK11 inactivation in liver-specific signaling and subsequent cancer growth and immune evasion mechanisms. This protocol describes in vitro validation of the downstream effects of STK11-AMPK signaling on HNF1A and FGL1 in a liver and two lung cancer cell lines.

The associated publication reports proteogenomic analysis of non-small cell lung cancer, where we identified molecular subtypes with distinct immune evasion mechanisms and therapeutic targets, and validated our classification method in separate clinical cohorts. This protocol describes the sample preparation and mass spectrometry (MS)-based in-depth and rapid proteomic analyses of tumor and biopsy samples. We deployed single-pot solid-phase-enhanced sample preparation (SP3). For the in-depth analysis, we used TMT labeling, followed by high-resolution isoelectric focusing (HiRIEF) prefractionation and LC-MS with data-dependent acquisition (DDA). The reported protocol achieved analytical depth of close to 14,000 quantified proteins and almost 10,000 across the entire cohort of 141 samples. The rapid analysis was label-free, based on LC-MS with data-independent acquisition (DIA). The median number of identified proteins was 3,967 and 3,552 in two independent cohorts of tumor samples (n = 141 and 208, respectively), and 2,494 in another cohort of biopsy material (n = 84).

Despite major advancements in lung cancer treatment, long-term survival is still rare and a deeper understanding of molecular phenotypes would allow the identification of specific cancer dependencies and immune-evasion mechanisms. Here we performed in-depth mass-spectrometry-based proteogenomic analysis of 141 tumors representing all major histologies of non-small cell lung cancer (NSCLC). We identified six distinct proteome subtypes with striking differences in immune cell composition and subtype-specific expression of immune checkpoints. Unexpectedly, high neoantigen burden was linked to global hypomethylation and complex neoantigens mapped to genomic regions, such as endogenous retroviral elements and introns, in immune-cold subtypes. Further, we linked immune evasion with LAG-3 via STK11 mutation-dependent HNF1A activation and FGL1 expression. Finally, we develop a data-independent acquisition mass-spectrometry-based NSCLC subtype classification method, validate it in an independent cohort of 208 NSCLC cases and demonstrate its clinical utility by analyzing an additional cohort of 84 late-stage NSCLC biopsy samples. Lehtio and colleagues perform proteogenomic analysis of non-small cell lung cancer and identify molecular subtypes with distinct immune-evasion mechanisms and therapeutic targets and validate their classification method in separate clinical cohorts.

BACKGROUND: Fibroblast Growth Factor 21 (FGF21) is expressed in several metabolically active tissues, including liver, fat and acinar pancreas, and has pleiotropic effects on metabolic homeostasis. The dominant source of FGF21 in the circulation is the liver. OBJECTIVE AND METHODS: To analyze the physiological functions of hepatic FGF21, we generated a hepatocyte specific knockout model (LKO) by mating albumin-Cre mice with FGF21 flox/flox (fl/fl) mice and challenged it with different nutritional models. RESULTS: Mice fed a ketogenic diet typically show increased energy expenditure; this effect was attenuated in LKO mice. LKO on KD also developed hepatic pathology and altered hepatic lipid homeostasis. When evaluated using hyperinsulinemic euglycemic clamps, glucose infusion rates, hepatic glucose production and glucose uptake were similar between fl/fl and LKO DIO mice. CONCLUSIONS: We conclude that liver derived FGF21 is important for complete adaptation to ketosis but has a more limited role in the regulation of glycemic homeostasis.

We report a rare case of a patient with AL amyloidosis and pseudoxanthoma elasticum skin lesions. An association between these two diseases has been previously described as amyloid elastosis in only six cases, but cardiac findings were not fully elucidated. The peculiarity of our case is that a severe cardiac involvement influenced the prognosis negatively. Furthermore, the electron microscopic examination did not show all the peculiar histopathological findings of amyloid elastosis, precluding a final diagnosis of this disease.

In this work, the authors will discuss about a new protocol and rapid alternative to isolate the mesenchymal stromal cells from the stromal vascular fraction, without the use of collagenase, to regenerate the bone tissue in the maxillo-facial region. This method employs a device that allows the isolation and concentration of stromal vascular fraction by means of lipoaspirates, which separate the lipid component and fragments of the extracellular matrix. The innovative element consists in using a filtration device instead of a centrifuging device to separate the different components. The purpose of this work was to illustrate the results obtained with the above-mentioned method in a series of 8 patients suffering from cystic neoformations maxillary or mandibular.

Pleomorphic adenoma is a common benign salivary gland tumor, which represents about 66 % of benign neoplasms of the salivary glands. Although it can occur in any salivary gland, it is most frequently found in the parotid. Pleomorphic adenomas are renowned for their cytomorphological and architectural heterogeneity that are characterized by intermixed epithelial and mesenchymal-like components. We report a rare case of pleomorphic adenoma of the parotid gland with prevalent schwannoma-like features mimicking a benign schwannoma. Microscopically the tumor showed a prevalence (about 95 %) of schwannoma-like areas with focal (about 5 %) epithelial component with tubular organization. The tumor showed positive immunoexpression for cytokeratin, S-100 protein, and focal expression of p63, CD10 and smooth muscle actin. To the best of our knowledge only six cases of schwannoma-like pleomorphic adenoma have been reported in the literature. The differential diagnosis between this entity and neurogenic and myogenic tumors is discussed.

Background The aim of the present study was to evaluate the prognostic significance of perineural invasion (PNI) in locally advanced gastric cancer patients who underwent D2 gastrectomy and adjuvant chemotherapy. Methods The records of a series of 103 patients undergoing D2 gastrectomy with curative intent combined with adjuvant chemotherapy from January 2004 to December 2014 were retrospectively reviewed. Results PNI was positive in 47 (45.6%) specimens. The 1-, 3-, and 5-year overall survival rates were 81%, 55%, and 42%, respectively. The 1-, 3-, and 5-year disease-free survival (DFS) rates were 76%, 57%, and 49%, respectively. A multivariate analysis showed that age number of positive lymph nodes, T stage, and PNI were independently associated with overall survival. Regarding DFS, the multivariate analysis showed that only PNI was independently associated with DFS. Conclusions PNI and T stage and positive lymph nodes are independent markers of poor prognosis in patients with gastric cancer. PNI should be incorporated in the postoperative staging system for planning follow-up after surgery and in our opinion to propose more aggressive postoperative therapies in PNI-positive patients.

With the widespread availability of biological antitumor drugs, the current scene of chemotherapies is changing. New chemotherapy agents, such as crizotinib, an inhibitor of anaplastic lymphoma kinase (ALK) and ROS1, usually used in pretreated advanced ALK-positive non-small-cell lung carcinoma, are more often used, and a description of the onset of side effects with suggestions for their management could be of interest for physicians. We describe a case of diffuse and aggressive renal polycystosis induced by crizotinib, which regressed after therapy, which could be of interest considering its wide extension and disappearance after the end of treatment. We also suggest some considerations from the literature and from the case reported that could be helpful in the management of this condition, which is known to be caused by crizotinib treatment.

Pure adenocarcinomas of the urothelium are very rare and their location in the pelvis is uncommon. Although their pathogenesis is not well defined, adenocarcinomas are likely to originate from neoplastic transformation of the glandular cells of the urothelial intestinal metaplasia usually arising in response to chronic irritating stimuli, such as long-duration inflammation, urolithiasis, and hydronephrosis. We report a case of an 81-year-old woman who underwent right nephrectomy for relapsing renal abscess due to a staghorn calculus. Histological examination disclosed an infiltrating adenocarcinoma arising from a tubulovillous adenoma with the surrounding pelvic mucosa showing a sequence of intestinal metaplasia, low- and high-grade villous adenoma, and invasive adenocarcinoma, supporting the hypothesis of cancer progression due to chronic inflammation from the urothelium through the metaplasia step.

Metastases of non-thyroid malignancies to the thyroid gland have been reported in 1.4% to 3% of patients undergoing thyroid surgery for thyroid malignancy. We report a case of thyroid metastases from renal cell carcinoma in a 57-year-old man, who underwent a left nephrectomy 11 years earlier for a renal cell carcinoma. The histological examination demonstrated a CD-10 positive and thyroglobulin and thyroid transcription factor-1 negative tissue, with numerous noncaseating gigantocellular granulomas. These findings are interesting for the possible role of the immune response in metastatic localizations.

We have previously demonstrated that expression of COX-2 is upregulated by hepatocyte growth factor in thyroid papillary carcinoma (TPC) cells and is associated with increased invasiveness of tumor cells. COX-2 upregulation was associated with downregulation of KAI-1/CD82, a metastasis suppressor molecule that has been associated with the metastatic potential of several solid tumors. In the present study, we have investigated the possibility that downregulation of KAI-1/CD82 may contribute to the invasiveness of papillary carcinoma cells.Expression of KAI-1/CD82 and its relation to COX-2 levels were investigated in 6 primary cultures of TPC, in 2 tumor cell lines (TPC-1 and K1), and in 55 tumor samples of TPC. The biological role of KAI-1/CD82 in regulating tumor invasiveness was investigated in TPC cell lines and primary cultures transfected with a pCDNA3.1/Hygro.KAI-1; transfected cells were tested in functional studies of cell migration and invasiveness. Finally, the role of KAI-1/CD82 in influencing TPC metastasis was investigated in vivo using nu/nu mice injected with K1-transfected cells.We provide evidence that COX-2 and KAI-1/CD82 are inversely regulated in TPC primary cultures and in TPC-1 tumor cells. In fact, inhibition of COX-2 with NS398 is associated with a 2-9-fold upregulation of KAI-1/CD82 RNA. Moreover, a possible relation between COX-2 and KAI-1/CD82 was confirmed by the presence of a statistically significant inverse correlation in the expression of the two genes in 55 tumor samples of TPC (r = -0.513; p = 0.001). In 36 of 55 cases, tumor areas contained lower levels of KAI-1/CD82 RNA as compared with the corresponding normal tissue. Low expression of KAI-1/CD82 RNA in the tumor area was associated with extrathyroid extension of the disease in 16 of 19 cases (p0.04) and with lymph node metastasis in 11 of 14 cases (not significant). KAI-1/CD82 re-expression in tumor cells was associated with a significant decrease in their migratory (50-76% reduction) and invasive (46-65% reduction) capacity, even after hepatocyte growth factor stimulation. Finally, nu/nu mice injected with KAI-1/CD82-transfected K1 TPC cells developed fewer and smaller metastasis as compared with mice injected with vector-transfected K1 cells (p=0.016).Our findings raise the possibility that downregulation of KAI-1/CD82 in TPC cells is one of the molecular mechanisms regulating their invasive and metastatic potential.