Current Options for Second-Line Systemic Therapy in Metastatic Renal Cell Carcinoma

Standard systemic therapy of advanced renal cell carcinoma (RCC) involves targeting angiogenesis, mainly through tyrosine kinase inhibitors (TKI) against the vascular endothelial growth factor receptor (VEGFR) pathway and targeting the immune checkpoints, namely, programmed death-1 (PD-1) or its ligand (PD-L1), and cytotoxic T-lymphocyte-associated protein 4 (CTLA4). With current strategies of combining these two approaches in the front-line setting, less is known about optimal selection of therapy upon development of resistance in the second and later lines of treatment for progressive disease. This review discusses currently available therapeutic options in patients who have progressive RCC after prior treatment with double immune check-point inhibitors (ICIs) or ICI-TKI combinations

Study of prognostic factors for urinary bladder cancer: investigation of NEP - neuropeptides and ubiquitin - proteasome - NFκB systems

The neutral endopeptidase (NEP/CD10)-neuropeptides (such as endothelin 1/ET-1) axis and the molecular pathway of the transcriptional nuclear factor kappaB (NFκB)-ubiquitin-proteasome are two mechanisms which have associated, independently from one another, with urinary bladder cancer. We studied these biological pathways as well as the relationship between them, using archived biological material (paraffin embedded tissue from transurethral resections of bladder tumors and radical cystectomies), in two groups of patients with bladder cancer. We prospectively studied patients with muscle-invasive bladder cancer and we retrospectively studied patients with non-muscle-invasive, high-grade bladder cancer. Finally, we performed immunohistochemical study for the following biological markers: NEP/CD10, ET-1 and NFκB (cytoplasmic).Our research hypothesis is that these two pathways are directly connected and interact in bladder cancer. Therefore, the neuropeptides could stimulate and activate the inactive cytoplasmic form of NFκB, resulting to its transfer into the nucleus (analogue mechanism of action with the extracellular stimulators of NFκB). As representative neuropeptide for testing our hypothesis we picked ET-1. If our hypothesis is correct, then the following should be proved: ↓ levels and/or activity of CD10 ► ↑ levels and activity of ΕΤ-1 ( ► ↑ cell proliferation, ↓ apoptosis and ↑ neovascularization ► negative clinical outcome in bladder cancer) ► ↑ proteasome activity ► ↑ proteasomal degradation IκB ► ↑ activity ΙΚΚ ► ↑ transcriptional activity of NF-κB ► ↑ NF-κB-dependent factors of angiogenesis, infiltration and metastasis ► negative clinical outcome in bladder cancer. Muscle-invasive bladder cancer patients:Only overexpression of ET-1 was significantly associated with negative clinical outcome, specifically with an increased hazard ratio for disease progression, in comparison with patients with mild expression of ET-1, throughout the duration of the monitoring period. Additionally, it was significantly associated with negative clinical outcome, specifically with an increased hazard ratio for death, in comparison with patients with moderate expression of ET-1, throughout the duration of the monitoring period. Finally, it was significantly associated with negative clinical outcome, specifically with an increased hazard ratio for death, in comparison with patients with mild expression of ET-1, throughout the duration of the monitoring period.Non-muscle invasive high grade bladder cancer patients:Expression of ET-1 was significantly associated with negative clinical outcome, specifically with an increased odds ratio for recurrence as well as progression of the disease and additionally with an increased hazard ratio for disease recurrence.Expression of NFκB was significantly associated with negative clinical outcome, specifically with an increased odds ratio for disease progression but not for recurrence.Muscle-invasive and non-muscle invasive high grade bladder cancer patients:Expression of NEP/CD10 was not associated with the studied prognostic parameters.Ο άξονας Ουδέτερης Ενδοπεπτιδάσης (NEP/CD10)-Νευροπεπτιδίων (όπως η ενδοθηλίνη 1/ET-1) και το βιολογικό μονοπάτι του μεταγραφικού πυρηνικού παράγοντα kappaB (NFκB)-Ουβικουϊτίνης-Πρωτεασώματος είναι δύο μηχανισμοί που έχουν συσχετιστεί, ανεξαρτήτως ο ένας από τον άλλον, με τον καρκίνο της ουροδόχου κύστης. Μελετήσαμε αυτές τις βιολογικές οδούς καθώς και τη μεταξύ τους σχέση, χρησιμοποιώντας αρχειακό βιολογικό υλικό (block παραφίνης από υλικό βιοψίας/εκτομής όγκων ουροδόχου κύστης ή κυστεκτομής), σε δύο ομάδες ασθενών με καρκίνο ουροδόχου κύστης. Μελετήσαμε προοπτικά ασθενείς με μυοδιηθητική νόσο και αναδρομικά ασθενείς με μη μυοδιηθητική, υψηλής κακοήθειας νόσο. Τέλος, πραγματοποιήσαμε ανοσοϊστοχημική μελέτη των μοριακών δεικτών NEP/CD10, ET-1 και NF-κB (κυτταροπλασματικό).Η ερευνητική υπόθεση είναι ότι οι δύο προαναφερθείσες οδοί είναι ευθέως και άμεσα συνδεδεμένες και αλληλεπιδρούν στον καρκίνο της ουροδόχου κύστης. Τα νευροπεπτίδια επομένως, μπορούν να δράσουν διεγερτικά και να ενεργοποιήσουν την ανενεργό κυτταροπλασματική μορφή του NF-κB, προκαλώντας μετακίνησή του εντός του κυτταρικού πυρήνα (ανάλογος μηχανισμός δράσης με τους εξωκυττάριους διεγέρτες του NF-κB). Ως αντιπροσωπευτικό νευροπεπτίδιο για την διερεύνηση της υπόθεσης επιλέξαμε την ενδοθηλίνη-1 (ΕΤ-1). Αν η υπόθεση είναι ορθή, τότε θα πρέπει να αποδειχθούν τα εξής:↓ επίπεδα και/ή δραστικότητα CD10 ► ↑ επίπεδα και δράση ΕΤ-1 ( ► ↑ κυτταρικού πολλαπλασιασμού, ↓ απόπτωσης και ↑ νεοαγγειογένεσης ► αρνητική κλινική έκβαση στον καρκίνο της ουροδόχου κύστης) ► ↑ δραστηριότητα πρωτεασώματος ► ↑ πρωτεασωμική αποδόμηση IκB ► ↑ δραστηριότητα ΙΚΚ ► ↑ μεταγραφική δραστηριότητα NF-κB ► ↑ NF-κB-εξαρτώμενων παραγόντων αγγειογένεσης, διείσδυσης και μεταστατικότητας ► αρνητική κλινική έκβαση στον καρκίνο της ουροδόχου κύστης. Στους ασθενείς με μυοδιηθητική νόσο:Μόνο η υπερέκφραση ΕΤ-1 συσχετίστηκε σε στατιστικώς σημαντικό βαθμό με αρνητική κλινική έκβαση, συγκεκριμένα με αυξημένη αναλογία κινδύνου (hazard ratio) για πρόοδο νόσου, σε σύγκριση με ασθενείς με ήπια έκφραση ΕΤ-1, καθόλη την διάρκεια της περιόδου παρακολούθησής τους. Επιπρόσθετα, συσχετίστηκε σε στατιστικώς σημαντικό βαθμό με αρνητική κλινική έκβαση, συγκεκριμένα με αυξημένη αναλογία κινδύνου (hazard ratio) για θάνατο, σε σύγκριση με ασθενείς με μέτρια έκφραση ΕΤ-1, καθόλη την διάρκεια της περιόδου παρακολούθησής τους. Τέλος, συσχετίστηκε σημαντικά με αρνητική κλινική έκβαση, συγκεκριμένα με αυξημένη αναλογία κινδύνου (hazard ratio) για θάνατο, σε σύγκριση με ασθενείς με ήπια έκφραση ΕΤ-1, καθόλη την διάρκεια της περιόδου παρακολούθησής τους.Στους ασθενείς με μη μυοδιηθητική, υψηλής κακοήθειας νόσο:Η έκφραση της ΕΤ-1 συσχετίστηκε σε στατιστικώς σημαντικό βαθμό με αρνητική κλινική έκβαση, συγκεκριμένα με αυξημένη αναλογία πιθανοτήτων (odds ratio) και για υποτροπή και για πρόοδο νόσου και επίσης με αυξημένη αναλογία κινδύνου (hazard ratio) για υποτροπή.Η έκφραση του NFκB συσχετίστηκε σε στατιστικώς σημαντικό βαθμό με αρνητική κλινική έκβαση, συγκεκριμένα με αυξημένη αναλογία πιθανοτήτων (odds ratio) για πρόοδο νόσου, αλλά όχι και για υποτροπή.Στους ασθενείς με μυοδιηθητική νόσο και τους ασθενείς με μη μυοδιηθητική, υψηλής κακοήθειας νόσο:Η έκφραση της NEP/CD10 δε συσχετίστηκε με τις υπο μελέτη προγνωστικές παραμέτρους

Radiofrequency-assisted, laparoscopic, clampless partial nephrectomy in patients with low-complexity small renal tumors: A retrospective cohort study

Background: This single-center, retrospective study was performed to investigate the safety and efficacy of radiofrequency-assisted (RF), laparoscopic partial nephrectomy (PN) with zero ischemia in patients with low-complexity small renal tumors. Materials and Methods: Patients with small renal masses (SRMs) who underwent laparoscopic, clampless laparoscopic partial nephrectomy - radiofrequency assisted (LPN-RFA) between January 2016 and June 2020 were studied. Demographics, clinical and pathological characteristics, recurrence-free survival, and overall survival were recorded. Results: Fifty-two SRMs were excised from corresponding patients using RFA–LPN. The median tumor size was 2.5 cm and all specimens involved low-complexity masses according to the renal nephrometry score. No conversions to radical nephrectomy were recorded. Postoperatively, there were one patient with fever, one with hematuria, and two with urinary leakage treated endoscopically. The majority of tumors (48/52, 86.2%) were clear-cell carcinomas. According to the glomerular filtration rate postoperatively and 12 months' posttreatment, adequate renal function was preserved in all patients. There were no positive surgical margins identified postoperatively and no recurrences during a median follow-up 24 months. All patients were alive at the last follow-up. Conclusions: This study suggests that RFA laparoscopic clampless PN represents an effective method for managing patients with low-complexity SRMs. It offers adequate intraoperative safety and excellent mid-term oncological control and functional preservation

Hypoxia-Inducible Factor-2-Altered Urothelial Carcinoma: Clinical and Genomic Features

Background: Hypoxia is recognized as a key feature of cancer growth and is involved in various cellular processes, including proliferation, angiogenesis, and immune surveillance. Besides hypoxia-inducible factor 1-alpha (HIF-1α), which is the main mediator of hypoxia effects and can also be activated under normoxic conditions, little is known about its counterpart, HIF-2. This study focused on investigating the clinical and molecular landscape of HIF-2-altered urothelial carcinoma (UC). Methods: Publicly available next-generation sequencing (NGS) data from muscle-invasive UC cell lines and patient tumor samples from the MSK/TCGA 2020 cohort (n = 476) were interrogated for the level of expression (mRNA, protein) and presence of mutations, copy number variations, structural variants in the EPAS1 gene encoding HIF-2, and findings among various clinical (stage, grade, progression-free and overall survival) and molecular (tumor mutational burden, enriched gene expression) parameters were compared between altered and unaltered tumors. Results: 19% (7/37) of UC cell lines and 7% (27/380) of patients with muscle-invasive UC display high EPAS1 mRNA and protein expression or/and EPAS1 alterations. EPAS1-altered tumors are associated with higher stage, grade, and lymph node metastasis as well as with shorter PFS (14 vs. 51 months, q = 0.01) and OS (15 vs. 55 months, q = 0.01). EPAS1 mRNA expression is directly correlated with that of its target-genes, including VEGF, FLT1, KDR, DLL4, CDH5, ANGPT1 (q EPAS1-altered tumors (9.9 vs. 4.9 mut/Mb), they are enriched in and associated with genes promoting immune evasion, including ARID5B, SPINT1, AAK1, CLIC3, SORT1, SASH1, and FGFR3, respectively (q Conclusions: HIF-2-altered UC has an aggressive clinical and a distinct genomic and immunogenomic profile enriched in angiogenesis- and immune evasion-promoting genes

Renin-Angiotensin System Single Nucleotide Polymorphisms Are Associated with Bladder Cancer Risk

The renin-angiotensin system (RAS), besides being a major regulator of blood pressure, is also involved in tumor angiogenesis. Emerging evidence suggests a correlation between the use of pharmacologic RAS inhibitors and a delay in urothelial bladder cancer (BC) progression. However, it is unknown whether RAS gene variants may predispose to the development of BC. This study examined the association of RAS single nucleotide polymorphisms (SNPs) including AT1R rs5186, AT2R rs11091046, REN rs12750834, ANG rs4762, and ANG rs699 with the risk of developing non-invasive BC. Peripheral blood samples from 73 patients with T1 urothelial BC (66 men, seven women) and an equal number of healthy subjects (control group) were collected. The TT genotype of the REN rs12750834 SNP (OR: 2.8 [1.3–6.05], p = 0.008) and to a lesser extent the presence of the T allele (OR: 2.3 [1.2–4.48], p = 0.01) conferred a higher risk of BC. The highest risk for BC within SNP carriers of the RAS system was associated with the presence of the CC genotype (OR: 17.6 [7.5–41.35], p p p < 0.001). In conclusion, these results support the clinical utility of RAS gene SNPs AT2R rs11091046, REN rs12750834, and ANG rs699 in the genetic cancer risk assessment of patients and families with BC

DNA library preparation for Ion Torrent S5 sequencing

The current protocol desrcibes the steps followed for DNA library preparation for NGS use from fresh-frozen kidney tissue samples, including cancerous and normal tissues. Library amplification, purification and quantification is included. </p

Endothelin-1 overexpression: a potential biomarker of unfavorable prognosis in non-metastatic muscle-invasive bladder cancer

Endothelin-1 (ET-1) is a multifunctional peptide exerting its effects via receptors A and B. ET-1 and its receptors, endothelin axis (ET axis), play a promoting role in cancer biology. Alterations of proteins of ET axis have been detected in non-metastatic muscle-invasive bladder cancer (NMMIBC). The objective of this study is to investigate the potential role of ET-1 tumor expression as a biomarker of prognosis, compared to other prognostic parameters (epidemiologic and pathologic), in NMMIBC. We prospectively included 40 consecutive, primary, high-grade NMMIBC patients. Tumor specimens after initial transurethral resection were stained immunohistochemically for ET-1. The ET-1 evaluation of expression was based on staining intensity (SI) of ET-1. SI was classified according to an arbitrary four-tiered scale (negative = 0, mild = 1, moderate = 2, strong = 3). Epidemiologic and pathologic parameters were analyzed, using univariate and multivariate statistics, for disease progression, progression-free survival (PFS), and overall survival (OS). ET-1 overexpression (SI = 3) was the unique parameter which associated significantly, both in univariate (log-rank test, p = 0.033) and multivariate (Cox regression analysis, p = 0.045, HR = 4.849, 95 % CI: 1.039-22.624) analysis, with an increased hazard ratio of progression. ET-1 overexpression (SI = 3) was also the unique parameter which associated, marginally significantly in univariate analysis (log-rank test, p = 0.056) and highly significantly in multivariate analysis (Cox regression analysis, p = 0.005, HR = 7.001, 95 % CI: 1.782-27.501), with an increased hazard ratio of death. Overexpression of ET-1 may be a potential biomarker of unfavorable prognosis in NMMIBC patients