Background and objective Failure rates after first-line treatment of localized prostate cancer (PCa) treatment remain high; therefore, it is essential to improve the selection and identification of at-risk patients to reduce mortality. The aim of the ANDROCAN study was to evaluate the biochemical recurrence (BCR) in patients with localized PCa treated by total prostatectomy at 5 yr after surgery, according to their presurgery gonadal status. Methods A prospective cohort study was conducted including 1318 patients undergoing total prostatectomy for localized PCa with a 5-yr postoperative follow-up. Clinical and hormonal data (assays of total testosterone [TT], bioavailable testosterone [BT], dihydrotestosterone, estrone, and estradiol were performed by gas chromatography/mass spectrometry) as well as metabolic syndrome parameters were collected at baseline before surgery. Pathological data (predominant Gleason grade 4 and stage) were collected and cross-referenced centrally. Factors associated with BCR were assessed by a multivariate analysis, and BCR-free survival was assessed by a Kaplan-Meier analysis. Key findings and limitations Among the 1318 patients, 237 had BCR of PCa. Considering demographic characteristics, populations with and without BCR were similar. However, patients with BCR had cancers with a higher Gleason score (p = 0.0001) and higher prostate-specific antigen (PSA) values (p = 0.0005) at baseline. Gleason score, pT >3a, and PSA level at baseline were positively correlated with BCR (p < 0.0001, p < 0.0001, and p = 0.0048, respectively), while BT and TT levels were not associated with BCR. This study includes patients with varying clinical characteristics, such as cancer history and metabolic syndrome, introducing variability that makes it challenging to isolate the specific effects of gonadal status on BCR. Another limitation is the lack of evaluation of long-term BCR beyond 5 yr, potentially overlooking recurrences that occur between 5 and 15 yr after surgery. This could lead to an underestimation of the actual long-term recurrence rates. Conclusions and clinical implications Overall, PSA levels, high Gleason score, and pT >3a are associated with a greater likelihood of disease recurrence following initial treatment and could serve as important prognostic indicators for predicting the risk of BCR. In this prospective study, biochemical hypogonadism was not associated with a higher occurrence of BCR within 5 yr of prostatectomy. The biological gonadal status of preoperative patients could potentially be useful for therapeutic decisions but does not provide an indication for the oncological follow-up. Patient summary Five-year follow up of patients after surgery showed that there is no association between hypogonadism (low levels of total testosterone and bioavailable testosterone) and cancer recurrence. However, cancer recurrence seems to be more associated with aggressiveness of cancer at the time of detection.

Biomedical samples are commonly used for histological examination after their inclusion in paraffin (Formalin-Fixed Paraffin-Embedded (FFPE) biopsy). However, they provide minimal information about the interactions between different tissue components such as blood vessels, nerves, and cellular aggregates. Here we present a modified iDISCO tissue-clearing method that we term miDISCO+. miDISCO+ can be used for analyzing FFPE samples, requires the use of only one single FFPE sample, and allows to acquire a detailed image of the 3D tissue/organ architecture and the relevant cellular interactions. The addition of an antigen retrieval step to the protocol enables to use antibodies whose binding is sensitive to formalin fixation due to antigen masking. This method enabled us to detect CD20+ B cell follicles and to show that they are in close contact with TH+ sympathetic nerve fibers in FFPE biopsies of human palatine tonsils, and to detect CD20+ B cells in lung tumors and observe their 3D organization within tertiary lymphoid structures. Thus, miDISCO+ could be a potent tool for clinicians to refine diagnosis and to select optimal personalized treatment by giving an integrated 3D view of the tissue structures and cellular interactions in single FFPE samples. ### Competing Interest Statement The authors have declared no competing interest.

Supplementary Figures 1-5 from Profound Coordinated Alterations of Intratumoral NK Cell Phenotype and Function in Lung Carcinoma

PDF file - 171K, Frequencies of gain, or loss, according to ERCC1-IHC status

PDF file, 686K, Heat maps of genes of the proliferative cluster (cluster f) according to ATAD2 expression, smoking status-associated discrete variables and disease stage in the LG cohort.

Supplementary Figure 2 from Intratumoral Induction of CD103 Triggers Tumor-Specific CTL Function and CCR5-Dependent T-Cell Retention

T-cell treatment with saracatinib or infection with shRNA targeting Pxn does not alter cell viability and cell surface protein expression levels

Supplementary Figure 6 from Profound Coordinated Alterations of Intratumoral NK Cell Phenotype and Function in Lung Carcinoma

PDF file - 123K, CD62L- T cells represent the main T cell population in human lung tumors.

PDF file - 78K, Multivariate Cox proportional hazards analysis for overall survival in NSCLC patients.

PDF file - 139K, Selective presence of CD62L+ T cells in TLS.

XLS file - 14K, List of Top 29 miRNA differentially expressed between the two groups ERCC1 positive / negative used for patients hierarchical clustering

PDF file - 380K, Gene expression levels related to immune populations, TLS, Th-orientation, cytotoxicity, T-cell activation, immuno-suppression, inflammation and angiogenesis according to the high and low density of mature DC.

PDF file - 131K, Clinical characteristic of NSCLC patients with DC-Lamp High versus DC-Lamp Low tumors.

PDF file - 65KB, Appendix A5.

PDF file - 95K, Comparison of different methods used for the stratification of patients according to the density of mature DC, stromal CD8+ T cells or tumor nest CD8+ T cells.

Table S1. Demographic and clinical characteristics of the analyzed patients; Table S2. Antibodies and conditions used for the IHC studies; Table S3. List of immune-related genes analyzed by Low Density Array; Figure S1. Gating and data analysis strategy; Figure S2. CD4+ and CD8+ T-cell differentiation in ccRCC TIL, and autologous PBL and RIL; Figure S3. Gap statistics according to the possible number of clusters of TIL phenotype. Optimal cut-off according to firstSEmax method (R package: cluster) is displayed (dotted line); Figure S4. PCA analysis including RIL and TIL phenotype. TIL clusters are displayed; Figure S5. Tumor size according to TIL clusters; Figure S6. Percentages of CD4+TIL expressing AM and InR according to tumor clusters. C1, Cluster1; C2, Cluster2; C3, Cluster3; Figure S7. Percentages of CD8+TIL expressing AM and InR according to tumor clusters. C1, Cluster1; C2, Cluster2; C3, Cluster3; Figure S8. Clonality Index and frequency of top 15 clonotypes in CD8+PD-1+ TIL according to tumor clusters. C1, Cluster1; C2, Cluster2; C3, Cluster3; Figure S9. Corrected P values for the differential gene expression between TIL clusters; Figure S10. Correlation matrix including TLS-related genes and immune cells densities in Immune-activated and Immune-regulated tumors; Figure S11. Percentages of CD4+PBL expressing differentiation markers, AM and InR in healthy controls (HC) and ccRCC-bearing patients; Figure S12. Percentages of CD8+PBL expressing differentiation markers, AM and InR in HC and ccRCC-bearing patients; Figure S13. Percentages of CD4+PBL expressing differentiation markers, AM and InR according to PBL Clusters; Figure S14. Percentages of CD8+PBL expressing differentiation markers, AM and InR according to PBL Clusters.

PDF file, 44K, LitVAn significant terms for gene clusters f and i in the unsupervised analysis of gene expression.

PDF file - 191K, Antibodies used in the study.

PDF file - 151K, Expression on tumor-infiltrating T cells of molecules involved in cytotoxicity, activation, and Th1 orientation.