Diagnosis Of Testicular Germ Cell Tumors Research Articles

Testicular germ cell tumor survival among children, adolescents, and young adults: A population-based retrospective cohort study.

10035 Background: While testicular germ cell tumor (TGCT) overall survival (OS) exceeds 90%, some patient groups, such as adolescents, have inferior event free survival (EFS) outcomes. There is a paucity of granular, population-derived data about outcomes in children, adolescents, and young adults (CAYA) with TGCT, limiting a deeper understanding of the factors that impact their survival. Methods: All male CAYA (ages 11-21 years) in Ontario, Canada diagnosed with TGCT between 1992-2021 were identified using provincial cancer registries. Detailed disease and treatment characteristics were chart abstracted for ~2/3 of the cohort and determined through health administrative databases for the remainder. We assessed 5-year OS and EFS (first of death, recurrence/progression, or subsequent malignant neoplasm [SMN]) using Kaplan-Meier analysis and the log rank test. Follow-up started at TGCT diagnosis, except for survival after SMN or recurrence, where follow-up started at event date. Results: 748 TGCT patients were identified; 521 were chart abstracted and thus had EFS data available. Median age at diagnosis was 19 years (interquartile range:18-21) and 83.6% had non-seminoma histology. OS and EFS ± standard error (SE) were (94.7 ± 0.8 %, n=748) and (76.4 ± 1.9%, n=521), respectively. Among patients with chart abstracted data, OS and EFS differed by cancer extent at diagnosis and were lowest among CAYA with non-lung organ metastases (Table). Patients who underwent retroperitoneal lymph node dissection (RPLND) for initial treatment (91/521) had higher survival than those who did not (OS 96.6 ± 1.9% vs 93.7 ± 1.2%, p=0.04; EFS 87.6 ± 3.5 % vs 74.1 ± 2.1 %, p=0.0008), particularly CAYA with lung-only organ metastases. OS post SMN (n=28) was 77.3 ± 8.2%, with lower OS after non-TGCT SMN (50.8 ± 14.4%, n=13) than second primary TGCT (100 ± 0%, n=15), p=0.002. OS after recurrence/progression (n=116) was superior in the 59 who did not receive chemotherapy at initial diagnosis (89.7 ± 4%) compared to the 57 who had initial chemotherapy (65.9 ± 6.4%; p=0.0001). Conclusions: CAYA with TGCT have low EFS, consistent with previous studies. The extent of metastasis is a significant predictor. Efforts are needed to improve the survival outcomes of young TGCT patients with organ metastasis at diagnosis, and those who recur or develop a non-TGCT SMN. [Table: see text]

Long-term outcomes of adolescents and young adults with testicular germ cell tumors: A population-based retrospective matched cohort study.

12016 Background: Testicular germ cell tumor (TGCT) survival exceeds 90%, but survivors of adult TGCT are at risk of developing late effects. Little is known about the risk for late effects in survivors of TGCT in children, adolescents, or young adults (CAYA). Methods: We identified all CAYA aged 11-21 years at TGCT diagnosis in Ontario, Canada, between 1992-2021. Those with complete treatment records were matched 1:5 by birth date and postal code at diagnosis to individuals from the general population (controls). Controls’ index date was defined as the diagnosis date of their survivor match. Participants were linked to health administrative databases to identify second malignant neoplasms (SMN); cardiovascular disease (CVD: congestive heart failure [CHF], myocardial infarction, coronary artery disease, pericardial disease, valvular abnormalities, arrhythmia, stroke, hypertension); renal disease (dialysis, kidney transplant); and hearing loss (physician-diagnosed hearing loss, hearing aid use). Cumulative incidences were compared between survivors and controls, and between chemotherapy and surgery treated survivors using Gray’s test. Cox proportional hazard models compared the risk of late effects between survivors and controls. Results: 521 TGCT survivors (296 chemotherapy treated, 225 treated with surgery only) were matched to 2605 controls. Median age at diagnosis/index date was 19 years. Survivors’ median follow-up time was 14.8 years (interquartile range: 10.7-21.7). Compared to controls, survivors had higher 15-year incidence and hazard ratio (HR) of all SMN (5.0% vs 0.5%; HR 8.4, p < .0001), non-testicular SMN (2.1% vs 0.5%; HR 3.8, p = 0.0003), dialysis (1.0% vs 0.04%; HR 12.4, p = 0.003), kidney transplant (0.4% vs 0.0%, p = 0.002), hearing loss (7.1% vs 3.1%; HR 1.8, p = 0.005), and hearing aid use (1.0% vs 0.2%; HR 4.1, p = 0.02). Survivors were at substantially increased risk of any CVD (8.2% vs 5.5%; HR 1.4, p < 0.03), CHF (1.2% vs. 0.2%; HR 5.0, p = 0.006), arrhythmia (1.8% vs 0.5%; HR 3.4, p = 0.002), and stroke (0.8% vs 0.05%; HR 8.7, p = 0.0006) despite no anthracycline exposure. Compared to surgery only treated survivors, chemotherapy treated survivors had higher 15-year incidence of any CVD (13.1% vs 2.0%, p < 0.0001), CHF (2.1% vs 0.0%, p = 0.03), arrhythmia (3.3% vs 0.0%, p = 0.02), stroke (1.4% vs 0.0%, p = 0.02), hypertension (7.4% vs 2.0%, p = 0.009), and hearing loss (10.2% vs 3.0%, p = 0.02). Conclusions: Survivors of TGCT during CAYA are at considerable risk for late effects affecting multiple organ systems. In addition to previously described morbidities like hearing loss, survivors (particularly those who received chemotherapy) are at higher risk of CVD. Despite excellent rates of cure, TGCT survivors diagnosed at a young age require life-long risk-directed follow-up to maximize their long-term health.

The values of systemic immune-inflammation index and neutrophil-lymphocyte ratio in predicting testicular germ cell tumors: A retrospective clinical study.

PurposeTo determine whether complete blood count (CBC) based inflammatory parameters can be used as markers predicting testicular germ cell tumors (TGCT).Material and methodsBetween 2013 to 2018 the data of 58 patients with testicular TGCT undergoing radical orchiectomy and 54 malignancy-free healthy men were retrospectively analyzed as tumor group and control group. Patient baseline characteristics including age, pathological stage and pre-surgery CBC based inflammatory parameters including neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), lymphocyte/monocyte ratio (LMR), systemic immune-inflammation index (SII), lymphocyte ratio (LR), neutrophil ratio (NR), mean platelet volume (MPV) and red cell distribution width (RDW) were analyzed and compared between tumor group and control group. Receiver operating characteristic (ROC) curve were used analyzing data with significantly difference to assess the discriminative ability of the markers for TGCT, area under the curve (AUC), cut-off value, sensitivity and specificity were calculated. The binary logistic regression model was used to evaluate the association between significant inflammatory markers and risk of TGCT.ResultsMean age of the tumor and control group was 41.1 ± 15.36 and 44.89 ± 9.2 years, respectively. Mean NLR, SII and RDW were significantly higher in tumor group compared with control group with P=0.005, P=0.001 and P=0.016, respectively; there were no significantly differences of age, PLR, LMR, LR, NR, MPV and RDW between groups. The ROC curve for NLR, SII and RDW was plotted in the diagnosis of TGCT and tumor progression, the cut-off value for NLR, SII and RDW were found as 3.38 (AUC: 0.704, sensitivity=51.4%, specificity=88.6%, P=0.003), 881.24 (AUC: 0.725, sensitivity=45.7%, specificity=91.4%, P=0.001) and 0.14 (AUC: 0.63, sensitivity=28.6%, specificity=97%, P=0.063), respectively. Patients were divided into two groups according to the threshold values, respectively. By using the multivariable logistic regression models, NLR ≥ 3.38 (OR, 5.86; 95% CI, 1.67-20.65, P=0.006) and SII ≥ 881.24 (OR, 4.89; 95% CI, 1.48-15.32, P=0.009) were independent risk factors predicting TGCT. Significantly statistical difference of pathological stage was also found between groups with respect to NLR cut-off values (P=0.034) and SII cut-off values (P=0.049). Combined the data together, NLR and SII both exhibited good differential diagnosis potential which could be used as markers predicting the TGCT.ConclusionAs the CBC based inflammation parameters, both NLR and SII could be used as effective tumor markers predicting the TGCT, and higher NLR and SII are associated with higher pathological stage. In addition, SII is a more powerful tool among these two inflammatory markers.

Long Non-Coding RNA RFPL3S Functions as a Biomarker of Prognostic and Immunotherapeutic Prediction in Testicular Germ Cell Tumor.

The incidence of testicular germ cell tumor (TGCT) is currently on the rise worldwide, of which 15%-30% of patients have occur recurrence and metastasis. However, clinical methods for diagnosing TGCT and judging its prognosis remained inadequate. In this study, we aimed to explore the possibility of testis-specific long-chain non-coding RNA (lncRNA) Ret finger protein-like 3S (RFPL3S) as a biomarker for TGCT diagnosis, prognosis, and treatment response by reviewing the TGCT gene expression data in Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. The cohort data and DNA methylation data of TGCT in TCGA were downloaded from TGCA, UCSC XENA, and GEO. The bioinformatic tools were used, including GEPIA2, Kaplan-Meier Plotter, LinkedOmics, UCSC XENA, Sangerbox Tools, GSCA, and Tumor Immune Dysfunction and Exclusion. Compared with normal testicular tissues, the RFPL3S expression was significantly reduced in TGCT, and was significantly negatively correlated with the patient’s Tumor, Node, Metastasis stage. Hypermethylation and low copy number of RFPL3S were present in TGCT, and low RFPL3S was associated with short disease-free and progression-free intervals. Silencing RFPL3S significantly enhanced the invasion ability and proliferation ability of TGCT cells as evaluated by Transwell and CCK-8 experiments. Additionally, RFPL3S expression was positively correlated with the infiltration of immune-activating cells such as B cells, CD8+ T cells, cytotoxic T cells, and natural killer cells, and negatively correlated with the infiltration of immunosuppressive cells such as Th17 and Th2. Higher RFPL3S expression was present in patients with immunotherapy benefits. In conclusion, we determined that the testis-specific lncRNA RFPL3S functioned as a tumor suppressor in TGCT and could be used as a prognostic predictor of TGCT, as well as a marker to predict the effect of TGCT immunotherapy.

Combining Hypermethylated RASSF1A Detection Using ddPCR with miR-371a-3p Testing: An Improved Panel of Liquid Biopsy Biomarkers for Testicular Germ Cell Tumor Patients.

Simple SummaryTesticular germ cell tumors are the most common solid cancers in men aged between 15–39 years. There is a need of non-invasive biomarkers for diagnosis and follow-up of these patients. miR-371a-3p has emerged as the most reliable biomarker of this disease, but fails to detect the teratoma subtype, which has clinical implications. In this work we describe a new method that combines miR-371a-3p quantification using RT-qPCR and hypermethylated RASSF1A quantification by droplet digital PCR in serum samples of these patients. The combination of both biomarkers detected disease (including teratoma) with a sensitivity of 100%, using cutoffs that made all healthy participants negative, being of interest for implementation in the clinical setting. The classical serum tumor markers used routinely in the management of testicular germ cell tumor (TGCT) patients—alpha fetoprotein (AFP) and human chorionic gonadotropin (HCG)—show important limitations. miR-371a-3p is the most recent promising biomarker for TGCTs, but it is not sufficiently informative for detection of teratoma, which is therapeutically relevant. We aimed to test the feasibility of hypermethylated RASSF1A (RASSF1AM) detected in circulating cell-free DNA as a non-invasive diagnostic marker of testicular germ cell tumors, combined with miR-371a-3p. A total of 109 serum samples of patients and 29 sera of healthy young adult males were included, along with representative cell lines and tumor tissue samples. We describe a novel droplet digital polymerase chain reaction (ddPCR) method for quantitatively assessing RASSF1AM in liquid biopsies. Both miR-371a-3p (sensitivity = 85.7%) and RASSF1AM (sensitivity = 86.7%) outperformed the combination of AFP and HCG (sensitivity = 65.5%) for TGCT diagnosis. RASSF1AM detected 88% of teratomas. In this representative cohort, 14 cases were negative for miR-371a-3p, all of which were detected by RASSF1AM, resulting in a combined sensitivity of 100%. We have described a highly sensitive and specific panel of biomarkers for TGCT patients, to be validated in the context of patient follow-up and detection of minimal residual disease.

Testicular Germ Cell Tumors: Classification, Pathologic Features, Imaging Findings, and Management

Testicular germ cell tumors (TGCTs) demonstrate a wide variety of histopathologic, genetic, pathogenetic, and immunocytochemical characteristics and various clinical-biologic profiles and prognoses. Most TGCTs arise from an intratubular precursor cell referred to as germ cell neoplasia in situ (GCNIS), which is an embryonic germ cell with the potential to differentiate into a plethora of embryonic and extraembryonic lineages. Advances in pathologic examination and genetics paved the way for the 2016 World Health Organization (WHO) classification system, which recognizes two pathogenetically distinct groups of TGCTs. Although postpubertal tumors originate from GCNIS, almost all prepubertal tumors belong to the non-GCNIS category. Molecular testing for chromosome 12p amplification helps to distinguish the two tumor categories. Imaging techniques such as US, CT, MRI, and fluorine 18 (18F)-fluorodeoxyglucose PET/CT are pivotal to the diagnosis and staging, evaluation of complications and treatment response, and long-term surveillance of TGCTs. In addition, select MRI findings may help to differentiate a seminoma from a nonseminomatous mixed TGCT. Accurate diagnosis of TGCTs has therapeutic and prognostic implications. Although seminomas show exquisite response to chemotherapy and radiation therapy, postpubertal teratomas are highly resistant to both. The 2016 WHO classification system introduced changes in the diagnosis and management of TGCTs, including the development of new treatment and follow-up guidelines. Radiologists play an essential role in the optimal treatment of patients with TGCTs. Online supplemental material is available for this article. ©RSNA, 2021.

Small RNAs in Seminal Plasma as Novel Biomarkers for Germ Cell Tumors.

Simple SummaryTesticular cancer is the most common cancer among young men. It is rarely diagnosed at early stages, being only detected with a highly invasive procedure that presents notable side-effects. Circulating small RNAs have recently been identified as testicular tumor markers, but are unable to diagnose testicular cancer at an early pre-invasive stage. So far, studies have been limited to microRNAs, with other small RNAs remaining unexplored as likely biomarkers. By sequencing all small RNAs in semen samples from men with different stages of testicular cancer and healthy men, we identify signatures predictive of cancer, even at an early stage. Thus, our study provides great potential for non-invasive early diagnosis of testicular cancer. Extensive biological variance in small RNA levels across samples, together with small sample sizes, limit the power to detect single small RNA markers. Hence, larger studies are needed to confirm our findings and deduce their full diagnostic capacity.Circulating miRNAs secreted by testicular germ cell tumors (TGCT) show great potential as novel non-invasive biomarkers for diagnosis of TGCT. Seminal plasma (SP) represents a biofluid closer to the primary site. Here, we investigate whether small RNAs in SP can be used to diagnose men with TGCTs or the precursor lesions, germ cell neoplasia in situ (GCNIS). Small RNAs isolated from SP from men with TGCTs (n = 18), GCNIS-only (n = 5), and controls (n = 25) were sequenced. SP from men with TGCT/GCNIS (n = 37) and controls (n = 22) were used for validation by RT-qPCR. In general, piRNAs were found at lower levels in SP from men with TGCTs. Ten small RNAs were found at significantly (q-value < 0.05) different levels in SP from men with TGCT/GCNIS than controls. Random forests classification identified sets of small RNAs that could detect either TGCT/GCNIS or GCNIS-only with an area under the curve of 0.98 and 1 in ROC analyses, respectively. RT-qPCR validated hsa-miR-6782-5p to be present at 2.3-fold lower levels (p = 0.02) in the SP from men with TGCTs compared with controls. Small RNAs in SP show potential as novel biomarkers for diagnosing men with TGCT/GCNIS but validation in larger cohorts is needed.

LAPTM5 Plays a Key Role in the Diagnosis and Prognosis of Testicular Germ Cell Tumors.

Objective Testicular germ cell tumors (TGCT) are a serious malignant tumor with low early diagnosis rates and high mortality. Methods To investigate novel biomarkers to predict the diagnosis and prognosis of this cancer, bioinformatics analysis was used as an accurate, efficient, and economical method. Results Our study detected 39 upregulated and 589 downregulated differentially expressed genes (DEGs) using the GEO and TCGA databases. To identify the function of DEGs, GO functional analysis, three pathway analysis (KEGG, REACTOME, and PANTHER), and protein-protein interaction network were performed using the KOBAS website, as well as the String database. After a series of analyses in GEPIA and TIMER, including differential expression, we found one candidate gene related to the prognosis and diagnosis of TGCT. LAPTM5 was also associated with CD8+ T cell and PDCD1 expression, which suggests that it may affect immune infiltration. Conclusions LAPTM5 was identified as a hub gene, which could be used as a potential biomarker for TGCT diagnosis and prognosis.

Origin of Subsequent Malignant Neoplasms in Patients with History of Testicular Germ Cell Tumor.

Simple SummaryAlthough testicular germ cell tumor (TGCT) carries a high cure rate, some patients still die from it. We investigated the genetic landscape and cellular origins of cancers that develop later in life after treatment for TGCT and found evidence that a common progenitor cell might be responsible for both. This study shows the possible importance of stem-like cells in the development of cancer.Although genetic changes may be pivotal in the origin of cancer, cellular context is paramount. This is particularly relevant in a progenitor germ cell tumor and its differentiated mature teratoma counterpart when it concerns tumor heterogeneity and cancer dormancy in subsequent second malignancies (subsequent malignant neoplasms (SMNs)). From our tumor registry database, we identified 655 testicular germ cell tumor (TGCT) patients who developed SMNs between January 1990 and September 2018. Of the 113 solid organ SMNs, 42 had sufficient tumor tissue available for fluorescence in situ hybridization (FISH) analysis of isochromosome 12p [i(12p)]. We identified seven additional patients for targeted DNA and RNA sequencing of teratomas and adjacent somatic transformation. Finally, we established cell lines from freshly resected post-chemotherapy teratomas and evaluated the cells for stemness expression by flow cytometry and by the formation of teratomas in a xenograft model. In our cohort, SMNs comprising non-germ cell tumors occurred about 18 years after a diagnosis of TGCT. Of the 42 SMNs examined, 5 (12%) contained i(12p) and 16 (38%) had 12p gain. When comparing a teratoma and adjacent somatic transformation, targeted DNA and RNA sequencing demonstrated high concordance. Studies of post-chemotherapy teratoma-derived cell lines revealed cancer-initiating cells expressing multipotency as well as early differentiation markers. For the first time, we demonstrated the prevalence of i(12p) in SMNs and the presence of progenitor cells embedded within mature teratomas after chemotherapy. Our findings suggest a progenitor stem-like cell of origin in SMN and TGCT and highlight the importance of cellular context in this disease.

Serum RNA Profiling in the 10-Years Period Prior to Diagnosis of Testicular Germ Cell Tumor.

Although testicular germ cell tumor (TGCT) overall is highly curable, patients may experience late effects after treatment. An increased understanding of the mechanisms behind the development of TGCT may pave the way for better outcome for patients. To elucidate molecular changes prior to TGCT diagnosis we sequenced small RNAs in serum from 69 patients who were later diagnosed with TGCT and 111 matched controls. The deep RNA profiles, with on average 18 million sequences per sample, comprised of nine classes of RNA, including microRNA. We found that circulating RNA signals differed significantly between cases and controls regardless of time to diagnosis. Different levels of TSIX related to X-chromosome inactivation and TEX101 involved in spermatozoa production are among the interesting findings. The RNA signals differed between seminoma and non-seminoma TGCT subtypes, with seminoma cases showing lower levels of RNAs and non-seminoma cases showing higher levels of RNAs, compared with controls. The differentially expressed RNAs were typically associated with cancer related pathways. Our results indicate that circulating RNA profiles change during TGCT development according to histology and may be useful for early detection of this tumor type.

Preoperative Inflammation Markers and De Ritis Ratio in Predicting Clinical Presentation and Prognosis of Patients with Testicular Germ Cell Tumors.

To evaluate the importance of neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), platelet-to-lymphocyte ratio (PLR), neutrophil-to-monocyte ratio (NMR) and De Ritis ratio (DRR) in predicting clinical presentation and prognosis of patients with testicular germ cell tumors (TGCTs). Observational study. Antalya Training and Research Hospital Antalya, Turkey, from January 2009 to March 2020. The characteristics and the results of biochemical and pathological examinations of patients who underwent radical orchiectomy were recorded. NLR, LMR, PLR, NMR, and DRR were calculated. The relationship among inflammation markers and DRR and clinical presentation and prognosis of TGCT was evaluated. Data of 99 patients were eligible for the study. Median age was 32 (27-39) years. Average size of the tumor was 5 (2.7 - 7) cm. Average duration of follow-up was 35.4 (8-62)months. Higher NLR and lower LMR were significantly correlated with higher rates of advanced-stage cancer, metastasis, and retroperitoneal lymph node invasion (RPLNI)(p<0.05). Based on the optimal cut-off values, there was a significantly higher rate of S stage, RPLNI, and metastatic disease in the high NLR group (p<0.05). Kaplan-Meier survival analysis found a statistically significantly lower mean survival rate in the high NLR group (p<0.05). There was no statistically significant difference between the DRR groups in the above-mentioned parameters (p>0.05). Preoperative NLR can be used as an inexpensive and easily accessible marker to predict clinical presentation at diagnosis and mortality rates during follow-up of patients with TGCT. Preoperative LMR can also be associated with the clinical picture at the time of diagnosis of TGCT. Key Words: De ritis ratio, Lymphocyte-to-monocyte ratio, Neutrophil-to-lymphocyte ratio, Neutrophil-to-monocyte ratio, Platelet-to-lymphocyte ratio, Testicular germ cell tumor.

Long-term survival in patients with germ cell testicular cancer: A population-based competing-risks regression analysis

Incidence of secondary malignancies and cardiovascular diseases among testicular germ cell tumor (TGCT) survivors is higher compared to the general population. We sought to describe the rates of other-cancer (OCM), non-cancer related (NCRM), and cancer-specific mortality (CSM) among men with TGCT. Using the Surveillance, Epidemiology, and End Results (SEER) database, 31,330 patients with a primary diagnosis of TGCT between 1973 and 2009 were identified. The primary endpoints comprised of 15-year CSM, OCM, and NCRM rates. Survival rates were stratified according to histology (seminoma vs. non-seminoma), median age (<34 vs. ≥34 years old), and disease stage (localized vs. regional vs. distant). Competing-risks Poisson regression methodologies were performed. For seminoma patients, the rates of CSM at 15 years increased with advancing stage (0.4-12.6%; P < 0.001), but varies little with age. In contrast, the rates of OCM (0.4-7.9%) and NCRM (2.9-8.9%) at 15 years increased with advancing stage and age (all P < 0.001). For non-seminoma patients, the 15-year CSM rates increased with advancing stage and age (1.9-24.4%; all P < 0.001). For the same time point, the rates of OCM (0.3-11.4%) and NCRM (2.4-8.0%) also increased with age and stage (all P ≤ 0.001). The risk of dying from secondary malignancies or other causes significantly increases with advancing stage and age at diagnosis among TGCT survivors. Such information can help provide patients and physicians with better screening strategies, follow-up protocols, and mental preparedness for such undesirable effects.

Spermatocytic seminoma in a 92-year-old man: report of a case in which SALL4, a potential novel marker for testicular germ cell tumors, was useful for the diagnosis

Spermatocytic seminoma (SS) is a rare testicular germ cell tumor (TGCT), occurring exclusively in the testes. Diagnosis of SS is important because, unlike other testicular tumors, SS must be treated by orchiectomy alone without further adjuvant therapy owing to its inability to metastasize. However, an accurate diagnosis of SS is often difficult given its rarity and the lack of a reliable marker. We report the occurrence of SS in a 92-year-old man—one of the oldest SS patients reported thus far. The patient presented with a painful right scrotal mass, which had been present for 6 months. A computed tomography scan revealed a testicular mass. There was no significant increase in the serum levels of alpha-fetoprotein (AFP) or human chorionic gonadotropin (hCG). A right orchiectomy was performed, which showed a well-circumscribed white-gray mass. A histological examination revealed sheets of non-cohesive cells with little intervening stroma. The tumor consisted of small, medium, and large cells with frequent mitoses, which is suggestive of SS. Whereas common TGCT markers including AFP, hCG, CD30, and placental alkaline phosphatase were all negative, nuclear reactivity for SALL4, a new TGCT marker, and positivity for CD117 (c-kit) finally helped establish the diagnosis of SS. We conclude that SALL4 immunostaining can be helpful for the diagnosis of TGCT when tumors are negative for conventional TGCT markers.

Testicular microlithiasis predicts concurrent testicular germ cell tumors and intratubular germ cell neoplasia of unclassified type in adults

There is an increasing body of literature associating testicular microlithiasis (TM), a common finding on testicular ultrasound, with testicular germ cell tumor (TGCT) and intratubular germ cell neoplasia of unclassified type (ITGCNU). Determining these associations is pertinent both clinically and biologically. To the authors' knowledge, no previous systematic review or meta-analysis has been performed. A comprehensive systematic literature review was performed without language restrictions through July 2009 and included an exhaustive search of electronic databases and article references. Two reviewers extracted data independently. Studies were categorized according to the clinical context in which sonography was performed. The primary study outcomes were concurrent diagnoses of TGCT or ITGCNU, with TM. In addition, studies with prospective follow-up of patients with TM were reviewed. Thirty-three studies met inclusion criteria. TM was not associated with an increased risk of TGCT in asymptomatic men. However, in referral populations, TM was associated overall with a risk ratio of 8.5 (95% confidence interval [CI], 4.5-16.1; P<.001) for a concurrent diagnosis of TGCT and 10.5 (95% CI, 5.3-20.8; P<.0001) for ITGCNU. Seventeen observational studies were identified in which the interval development of TGCT in patients with TM was reported; however, the majority of those studies did not report the follow-up of a control arm and could not be summarized. In the presence of risk factors, TM was associated with a substantially elevated risk of a concurrent diagnosis of TGCT and ITGCNU. The authors suggest modifications to recently proposed guidelines for the management of TM.

Seminal expression of NY‐ESO‐1 and MAGE‐A4 as markers for the testicular cancer

Testicular germ cell tumours (TGCTs) are the most common malignancies in Caucasian young men and their incidence has increased over the past decades. However, a non-invasive test allowing an early diagnosis of TGCT often proves inaccurate. We have previously shown that two Cancer-Testis Antigens (CTA), namely MAGE-A4 and NY-ESO-1, were expressed by TGCT. As exfoliation of carcinoma in situ (CIS) cells or tumour germ cells from testis into seminal fluid can occur, here we studied the expression of the 2 CTA in semen smears of patients with testicular cancer in comparison with healthy men. Using semen smears from healthy controls (n = 65) and patients diagnosed for testicular tumour (n = 57) and immunological staining, we observed expression of MAGE-A4 and NY-ESO-1 proteins in seminal fluid exfoliated cells. We found a highly statistically significant difference in the ratios of stained cells to the total number of round cells between testicular cancer patients and healthy controls. Multivariable analysis, including sperm parameters and immunostaining on sperm smears, shows the improvement. This technique can provide towards testicular cancer diagnosis when it is included in the current testing regime. However, the fact that expression of these markers was not restricted to foetal germ cells led to detection in the semen of a number of healthy subjects. Although the detection of these CTA could be useful to characterize the sub-type of individual TGCTs better, we stress here that the false positive rate precludes the exclusive employment of these CTA for the early detection of testicular neoplasia.

Increasing incidence of testicular germ cell tumors among black men in the United States: McGlynn KA, Devesa SS, Graubard BI, Castle PE, Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Department of Health and Human Services, Rockville, MD

There has been marked disparity in the incidence of testicular germ cell tumors (TGCT) among white and black men for a number of decades in the United States. Since at least the beginning of the Surveillance, Epidemiology, and End Results (SEER) Program in 1973, incidence rates among white men have been five times higher than rates among black men. In addition, rates among white men have been increasing, whereas rates among black men have remained stable. However, a recent examination of ethnic-specific rates suggested that the incidence among black men may have begun to change in the 1990s. TGCT incidence data from nine registries of the SEER Program were analyzed for the years 1973 to 2001. Trends were examined separately for seminoma and nonseminoma. Analyses found that the incidence of TGCT began to increase among black men between the 1988 to 1992 and 1993 to 1997 periods. Before that time, incidence among black men had decreased by 14.8%. Between 1988 to 1992 and 1998 to 2001, however, the incidence increased by 100%, with the incidence of seminoma increasing twice as much (124.4%) as the incidence of nonseminoma (64.3%). Over the 29-year time period, there was no evidence of a change in the proportion of tumors diagnosed at earlier stages among black men. In contrast, the proportion of tumors diagnosed at localized stages significantly increased among white men. The incidence of TGCT among black men has increased since 1988 to 1992. Although the reasons for this increase are unclear, screening and earlier diagnosis of TGCT do not seem to be factors.

Increasing incidence of testicular germ cell tumors among black men in the United States.

There has been marked disparity in the incidence of testicular germ cell tumors (TGCT) among white and black men for a number of decades in the United States. Since at least the beginning of the Surveillance, Epidemiology, and End Results (SEER) Program in 1973, incidence rates among white men have been five times higher than rates among black men. In addition, rates among white men have been increasing, whereas rates among black men have remained stable. However, a recent examination of ethnic-specific rates suggested that the incidence among black men may have begun to change in the 1990s. TGCT incidence data from nine registries of the SEER Program were analyzed for the years 1973 to 2001. Trends were examined separately for seminoma and nonseminoma. Analyses found that the incidence of TGCT began to increase among black men between the 1988 to 1992 and 1993 to 1997 periods. Before that time, incidence among black men had decreased by 14.8%. Between 1988 to 1992 and 1998 to 2001, however, the incidence increased by 100%, with the incidence of seminoma increasing twice as much (124.4%) as the incidence of nonseminoma (64.3%). Over the 29-year time period, there was no evidence of a change in the proportion of tumors diagnosed at earlier stages among black men. In contrast, the proportion of tumors diagnosed at localized stages significantly increased among white men. The incidence of TGCT among black men has increased since 1988 to 1992. Although the reasons for this increase are unclear, screening and earlier diagnosis of TGCT do not seem to be factors.

Immunohistochemical heterogeneity of type 1 blood group antigen expressions in testicular germ cell tumors.

The immunohistochemical expression of type 1 blood group antigens (type 1 BGAs) was analyzed for 30 cases of testicular germ cell tumors (TGCTs), using monoclonal antibodies against DU-PAN-2, CA19-9, Lewis(a) (Le(a)), and Lewis(b) (Le(b)). DU-PAN-2 was expressed very frequently in all of the embryonal carcinomas (ECs). CA19-9 expression was demonstrated in 53% of ECs, but the number of positive cells was generally smaller than that for DU-PAN-2. CA19-9-negative ECs tended to show a higher number of DU-PAN-2-positive cells compared to CA19-9-positive ECs, and ECs in which DU-PAN-2 was more strongly expressed showed a relatively frequent expression of CA19-9. In 36% of seminomas and 56% of yolk sac tumors (YSTs), DU-PAN-2 was weakly expressed, and the positive cells were few in number. Little or no expression of CA19-9 was demonstrated in seminomas and YSTs. Regarding Le(a) and Le(b), the expressions were found to be limited to teratomas at a frequency of 57% and 86%, respectively, with the exception of one EC positive for Lea and one YST positive for Leb. Eighty-six percent of teratomas showed expressions of DU-PAN-2 and CA19-9. DU-PAN-2 was also seen in some intratubular malignant germ cells. The antibodies used were all negative for choriocarcinomas, syncytiotrophoblastic giant cells, and normal testicular tissues. The antigen expressions were predominantly observed on the surface of tumor cells developing luminal structures. In conclusion, although CA19-9 was relatively specific for ECs, it should be emphasized that ECs were rather characteristic of extensive DU-PAN-2 expression. Particularly in CA19-9-negative ECs, a combined analysis of DU-PAN-2 and CA19-9 would be helpful in confirming the histopathologic diagnosis of TGCTs. The clinical significance of DU-PAN-2 in ECs as a tumor marker remains to be clarified. Le(a) and Le(b) expressions were thought to be related to the differentiation or maturation rather than to the malignant transformation in TGCTs.

Heterogeneity in alkaline phosphatase isozyme expression in human testicular germ cell tumours: An enzyme-/immunohistochemical and molecular analysis

In humans, alkaline phosphatases are encoded by one tissue-non-specific alkaline phosphatase (TNAP) gene and three tissue-specific alkaline phosphatase genes, intestinal, placental (PLAP), and germ cell-specific alkaline phosphatase (GCAP). Although the presence of alkaline phosphatases in testicular germ cell tumours (TGCTs) of adolescents and adults has been utilized for both detection and patient monitoring, it is not known in detail which isozymes are expressed. Since alkaline phosphatase is detected in carcinoma in situ (CIS), the common precursor of all TGCTs, it might provide a marker for the early diagnosis of TGCTs. Testicular cancers of germ cell and non-germ cell origin along with testicular parenchyma with and without CIS have been analysed for the expression of the different alkaline phosphatase isozymes. Antibodies to TNAP and PLAP/GCAP showed positivity in CIS, seminoma, and embryonal carcinoma. The heterogeneous staining pattern detected in frozen tissue sections was similar to the pattern found in formalin-fixed, paraffin-embedded material, indicating a biological phenomenon and not a handling artefact. Since PLAP and GCAP cannot be distinguished using immunohistochemistry, the expression of these isozymes was studied at the molecular level using a reverse transcriptase-polymerase chain reaction (RT-PCR) approach, in combination with a primer extension assay. The results show that CIS and seminoma predominantly express GCAP, while in embryonal carcinoma the expression of GCAP versus PLAP varies. Due to the presence of alkaline phosphatase transcripts in normal testicular parenchyma, an RT-PCR-based analysis of alkaline phosphatase is not informative for the early detection of TGCTs in biopsy samples.

Detection of Human Endogenous Retrovirus Type K-Specific Transcripts in Testicular Parenchyma and Testicular Germ Cell Tumors of Adolescents and Adults: Clinical and Biological Implications

Testicular germ cell tumors (TGCTs) of adolescents and adults have been shown to contain proteins of the human endogenous retrovirus type K family. In a recent study, expression of these retroviral sequences was confirmed using in situ hybridization, which also showed expression in carcinoma in situ, the precursor of all TGCTs. Because of the clinical significance of a test for early diagnosis of TGCTs, we studied whether expression of human endogenous retrovirus type K genes could be an informative parameter. Therefore, we investigated TGCTs of various histologies and testicular parenchyma with and without carcinoma in situ using reverse transcription-polymerase chain reaction for expression of the gag, env, and prt genes. The gag and prt genes were expressed in all samples tested. The env transcripts were not found in TGCTs showing somatic differentiation only but could be detected in most normal testicular parenchyma samples. Therefore, detection of human endogenous retrovirus type K transcripts cannot be used for early diagnosis of TGCTs. Simultaneous expression of multiple gag sequences was found both in normal parenchyma and TGCTs, and we demonstrated that expression of gag sequences with an extra G, necessary to generate a functional protein, was not limited to TGCTs.