Prognostic significance of programmed cell death-ligand 1 expression on circulating tumor cells in various cancers: A systematic review and meta-analysis.

BACKGROUND: The prognostic significance of programmed cell death-ligand 1 (PD-L1) expression on circulating tumor cells (CTCs) has been explored but is still in controversy. We performed, for the first time, a meta-analysis to systematically evaluate its prognostic value in human cancers. METHODS: Literature databases were searched for eligible studies prior to June 30, 2021. The pooled hazard ratios (HRs) and 95% confidence intervals (95% CIs) were calculated for the associations of pre-treatment and post-treatment PD-L1+ CTCs with progression-free survival (PFS) and overall survival (OS). Subgroup analyses with regards to cancer type, treatment, CTC enrichment method, PD-L1 detection method, cut-off, and specifically the comparison model were performed. RESULTS: We included 30 eligible studies (32 cohorts, 1419 cancer patients) in our analysis. Pre-treatment PD-L1+ CTCs detected by immunofluorescence (IF) tended to predict better PFS (HR = 0.55, 95% CI 0.28-1.08, p = 0.084) and OS (HR = 0.61, 95% CI 0.36-1.04, p = 0.067) for immune checkpoint inhibitor (ICI) treatment, but were significantly associated with unfavorable survival for non-ICI therapies (PFS: HR = 1.85, 95% CI 1.21-2.85, p = 0.005; OS: HR = 2.44, 95% CI 1.69-3.51, p < 0.001). Post-treatment PD-L1+ CTCs predicted markedly worse PFS and OS. The prognostic value was obviously modulated by comparison models. Among patients with detectable CTCs, PD-L1+ individuals had comparable survival to PD-L1- individuals, except ICI treatment for which PD-L1+ may predict better PFS (HR = 0.42, 95% CI 0.17-1.06, p = 0.067). Patients with PD-L1+ CTCs had worse survival prognosis compared to those without PD-L1+ CTCs in overall analysis (PFS: HR = 2.10, 95% CI 1.59-2.77, p < 0.001; OS: HR = 2.55, 95% CI 1.70-3.81, p < 0.001) and in most subgroups. CONCLUSIONS: Our analysis demonstrated that PD-L1 positive expression on CTCs predicted better survival prognosis for ICI treatment but worse survival for other therapies, which thus can be potentially used as a prognostic marker of malignant tumor treatment. However, the prognostic value of PD-L1+ CTCs for ICI treatment needs validation by more large-scale studies in the future.

An ionising radiation-induced specific transcriptional signature of inflammation-associated genes in whole blood from radiotherapy patients: a pilot study.

BACKGROUND: This communication reports the identification of a new panel of transcriptional changes in inflammation-associated genes observed in response to ionising radiation received by radiotherapy patients. METHODS: Peripheral blood samples were taken with ethical approval and informed consent from a total of 20 patients undergoing external beam radiotherapy for breast, lung, gastrointestinal or genitourinary tumours. Nanostring nCounter analysis of transcriptional changes was carried out in samples prior and 24 h post-delivery of the 1st radiotherapy fraction, just prior to the 5th or 6th fraction, and just before the last fraction. RESULTS: Statistical analysis with BRB-ArrayTools, GLM MANOVA and nSolver, revealed a radiation responsive panel of genes which varied by patient group (type of cancer) and with time since exposure (as an analogue for dose received), which may be useful as a biomarker of radiation response. CONCLUSION: Further validation in a wider group of patients is ongoing, together with work towards a full understanding of patient specific responses in support of personalised approaches to radiation medicine.

Comprehensive analysis of the effect of rs2295080 and rs2536 polymorphisms within the mTOR gene on cancer risk.

There is still no conclusion on the potential effect of the rs2295080 and rs2536 polymorphisms of mTOR (mammalian target of rapamycin) gene on different cancers. Herein, we performed a comprehensive assessment using pooled analysis, FPRP (false-positive report probability), TSA (trial sequential analysis), and eQTL (expression quantitative trait loci) analysis. Eighteen high-quality articles from China were enrolled. The pooled analysis of rs2295080 with 9502 cases and 10,965 controls showed a decreased risk of urinary system tumors and specific prostate cancers [TG vs. TT, TG+GG vs. TT and G vs. T; P<0.05, OR (odds ratio) <1]. FPRP and TSA data further confirmed these results. There was an increased risk of leukemia [G vs. T, GG vs. TT, and GG vs. TT+TG genotypes; P<0.05, OR>1]. The eQTL data showed a potential correlation between the rs2295080 and mTOR expression in whole blood samples. Nevertheless, FPRP and TSA data suggested that more evidence is required to confirm the potential role of rs2295080 in leukemia risk. The pooled analysis of rs2536 (6653 cases and 7025 controls) showed a significant association in the subgroup of "population-based" control source via the allele, heterozygote, dominant, and carrier comparisons (P<0.05, OR>1). In conclusion, the TG genotype of mTOR rs2295080 may be linked to reduced susceptibility to urinary system tumors or specific prostate cancers in Chinese patients. The currently data do not strongly support a role of rs2295080 in leukemia susceptibility. Large sample sizes are needed to confirm the potential role of rs2536 in more types of cancer.

Dicentric Dose Estimates for Patients Undergoing Radiotherapy in the RTGene Study to Assess Blood Dosimetric Models and the New Bayesian Method for Gradient Exposure.

The RTGene study was focused on the development and validation of new transcriptional biomarkers for prediction of individual radiotherapy patient responses to ionizing radiation. In parallel, for validation purposes, this study incorporated conventional biomarkers of radiation exposure, including the dicentric assay. Peripheral blood samples were taken with ethical approval and informed consent from a total of 20 patients undergoing external beam radiotherapy for breast, lung, gastrointestinal or genitourinary tumors. For the dicentric assay, two samples were taken from each patient: prior to radiotherapy and before the final fraction. Blood samples were set up using standard methods for the dicentric assay. All the baseline samples had dicentric frequencies consistent with the expected background for the normal population. For blood taken before the final fraction, all the samples displayed distributions of aberrations, which are indicative of partial-body exposures. Whole-body and partial-body cytogenetic doses were calculated with reference to a 250-kVp X-ray calibration curve and then compared to the dose to blood derived using two newly developed blood dosimetric models. Initial comparisons indicated that the relationship between these measures of dose appear very promising, with a correlation of 0.88 (P = 0.001). A new Bayesian zero-inflated Poisson finite mixture method was applied to the dicentric data, and partial-body dose estimates showed no significant difference (P > 0.999) from those calculated by the contaminated Poisson technique. The next step will be further development and validation in a larger patient group.

New plasma preparation approach to enrich metabolome coverage in untargeted metabolomics: plasma protein bound hydrophobic metabolite release with proteinase K.

Plasma untargeted metabolomics is a common method for evaluation of the mechanisms underlying human pathologies and identification of novel biomarkers. The plasma proteins provide the environment for transport of hydrophobic metabolites. The current sample preparation protocol relies on the immediate precipitation of proteins and thus leads to co-precipitation of a significant fraction of hydrophobic metabolites. Here we present a new simple procedure that overcomes the co-precipitation problem and improves metabolome coverage. Introducing an additional step preceding the protein precipitation, namely limited digestion with proteinase K, allows release of associated metabolites through the relaxation of the native proteins tertiary structure. The modified protocol allows clear detection of hydrophobic metabolites including fatty acids and phospholipids. Considering the potential involvement of the hydrophobic metabolites in human cardiovascular and cancer diseases, the method may constitute a novel approach in plasma untargeted metabolomics.

The effect of anaesthetic management on neutrophil gelatinase associated lipocalin (NGAL) levels after robotic surgical oncology.

PURPOSE: The main objective of this study was to compare the effect of two anaesthetic techniques (general vs combined) on plasma levels of NGAL (Neutrophil Gelatinase Associated Lipocalin) after robotic urogenital oncosurgery. The secondary objective was to correlate NGAL levels with the incidence of acute kidney injury (AKI). METHODS: This was a longitudinal prospective study. Forty patients were included and randomized in 2 groups: group C (N=16 cases; combined general-epidural anesthesia) and group G (N=24 cases; control group with general anesthesia). Demographic data, Charlson Comorbidity Index, Apache II, SOFA and ASA scores were similar in both groups. Serum creatinine was determined preoperatively and every 24 hrs for 4 postoperative days to identify AKI according to RIFLE and AKIN criteria. Serum NGAL was determined at 6 and 12 hrs after induction of anesthesia. RESULTS: Serum creatinine increased at 24 hrs postoperatively in both groups as compared to baseline, but significant changes were registered only in the G group (p(control) = 0.004). Serum NGAL increased significantly in both groups as compared with baseline levels (pcase=0.0034 vs p(control)=0.0001). The incidence of AKI was 12.50% (95% CI 0.4-34) in the C group and 37.50% (95% CI 17-58) in the G group (p=0.0909), respectively. CONCLUSION: Impaired renal function and AKI occurred in robot-assisted laparoscopic urogenital oncosurgery under both general and combined anaesthesia. The incidence of AKI was lower in patients undergoing combined anesthesia compared to general anaesthesia after robotic urogenital oncosurgery but the difference did not reach statistical significance. However, plasma levels of NGAL were significantly increased at 6 and 12 hrs in the general anaesthesia group as compared with combined anaesthesia. NGAL may be a better marker in detecting postoperative acute kidney injury. Further studies are needed.

Risk factors for refractory febrile neutropenia in urological chemotherapy.

During chemotherapy, patients are more susceptible to infectious complications as a result of bone marrow suppression, leading to neutropenia. The purpose of this study is to investigate risk factors for refractory febrile neutropenia (FN) during urological chemotherapy. Our method for suppressing FN is to use granulocyte colony-stimulating factor and prevent upper respiratory infection by masking and gargling. We studied 47 episodes of FN in 39 patients that occurred during urological chemotherapy for urothelial cancer, testicular cancer, and prostate cancer. Among our cases, there were 5 patients with refractory FN; we set risk factors for refractory FN and performed statistical analyses. The average age of the 39 patients was 60.6 years (range, 18-80 years). In 47 FN episodes, the chemotherapy regimen before the occurrence of FN included 15 (31.9 %) MVAC (methotrexate, vinblastine, adriamycin, cisplatin) for urothelial cancer, 5 (10.6 %) DE (docetaxel, estramustin) for prostate cancer, and 3 (6.4 %) TIP (paclitaxel, ifosfamide, cisplatin) for testicular cancer. The antibiotics used to treat FN included 17 (36.3 %) meropenem and 23 (49.0 %) cefepime, and the average duration of antibiotics was 4.4 days (range, 1-12). We investigated risk factors for refractory FN and showed a significant relationship between refractory FN and indwelling urinary catheter or smaller Multinational Association for Supportive Care in Cancer score by multivariate analysis. A future prospective study is needed for further evaluation for risk factors and establishing treatment protocols and guidelines for FN.

[Identification of circulating tumor cells as a promising method of genitourinary cancer diagnosis]. / Identyfikacja krazacych komórek nowotworowych jako obiecujaca metoda diagnostyki chorób nowotworowych ukladu moczowo-plciowego.

Circulating tumor cells (CTCs) are cells circulating in the blood, which in terms of antigenic or genetic profile correspond to a particular type of cancer. It is suspected that CTCs possess properties of cancer stem cells. Detection, quantification and characterization of CTCs in the peripheral blood can be of great importance for modern oncology. In the case of early-stage disease, CTCs may help in cancer detection, estimation of metastasis risk and treatment prognosis. In advanced cancer patients, CTCs may also have prognostic significance and may facilitate monitoring response to treatment. Identification of CTCs in the circulation and their differentiation from hematopoietic cells and normal epithelial cells could be based on physical and biological properties such as size, density and expression of specific proteins. Immunomagnetic techniques are the most commonly used methods of CTCs isolation. CellSearch System (CSS) is the only test for detecting CTCs in the peripheral blood approved by the Food and Drug Administration (FDA) for clinical use. The paper presents the characteristics of circulating tumor cell isolation methods and the results of studies concerning CTCs isolation in patients with prostate, bladder and kidney cancer. 

[Value of biomarkers in urology]. / Stellenwert von Biomarkern in der Urologie.

As part of the diagnosis and treatment of cancer objective biological measures are becoming increasingly important and may help to elicit the individual patient's risk of future outcome. Among theses measures are markers as determinants for the biological potential of the malignant disease, the prediction of recurrence after systemic treatment or the response to a specific therapy. In counselling patients with uro-oncologic diseases, the emerging role of evidence-based treatment choices reveals with cumulative certainty that the available information on markers is inconclusive.The aim of this review is to provide a summary of the current evidence of the evaluation of markers, which may be supportive for a treatment decision and/or provide additional valuable information as part of the treatment option. In addition, this may provide access to some of the most promising investigations, which may yield perspectives for future relevant clinical questions.

Immunohistochemical analysis of Reg IV in urogenital organs: Frequent expression of Reg IV in prostate cancer and potential utility as serum tumor marker.

Regenerating islet-derived family, member 4 (Reg IV) is a candidate marker for cancer and inflammatory bowel disease and is associated with neuroendocrine and intestinal differentiation. We have reported that 14% of prostate cancer (PCa) cases are positive for Reg IV by immunohistochemistry. In the present study, we performed immunohistochemical analysis of Reg IV in other major urological cancers, including 101 renal cell carcinoma (RCC), and 95 urothelial carcinoma (UC) of urinary bladder by immunohistochemistry. We also investigated neuroendocrine differentiation by chromogranin A and synaptophysin staining along with intestinal differentiation by MUC2 staining. Immunohistochemical analysis of Reg IV revealed no expression of Reg IV in RCC, and only one case (1%) of UC expressed Reg IV. Neither neuroendocrine nor intestinal differentiation was found in RCC. Among 95 UC cases, neuroendocrine differentiation was detected in 13 cases (14%), and intestinal differentiation was observed in 33 cases (35%). In one Reg IV-positive UC case, MUC2 staining was observed. Since Reg IV expression was frequently found in PCa, we also measured Reg IV levels in sera from patients with PCa by enzyme-linked immunosorbent assay. The serum Reg IV concentration in PCa patients (n=38, mean +/- SE, 1.69+/-0.16 ng/ml) was significantly higher than that in control individuals (n=40, 1.28+/-0.11 ng/ml, P=0.0199, Mann-Whitney U test). The sensitivity and specificity for detection of PCa were 34% (13/38) and 90% (36/40), respectively. These results suggest that among major urologic cancers, Reg IV is expressed frequently in PCa, and that serum Reg IV represents a novel biomarker for PCa.

Elevated rates of testosterone-related disorders in women with autism spectrum conditions.

The androgen theory of autism proposes that autism spectrum conditions (ASC) are in part due to elevated fetal testosterone (FT) levels, which are positively correlated with a number of autistic traits and inversely correlated with social development and empathy. A medical questionnaire was completed by n=54 women with ASC, n=74 mothers of children with ASC, and n=183 mothers of typically developing children to test whether women with ASC have an increased rate of testosterone-related medical conditions, and to see whether mothers of children with ASC show similar abnormalities, as part of the 'broader autism phenotype'. Compared to controls, significantly more women with ASC reported (a) hirsutism, (b) bisexuality or asexuality, (c) irregular menstrual cycle, (d) dysmenorrhea, (e) polycystic ovary syndrome, (f) severe acne, (g) epilepsy, (h) tomboyism, and (i) family history of ovarian, uterine, and prostate cancers, tumors, or growths. Compared to controls, significantly more mothers of ASC children reported (a) severe acne, (b) breast and uterine cancers, tumors, or growths, and (c) family history of ovarian and uterine cancers, tumors, or growths. These results suggest current hormone abnormalities in women with ASC and their mothers. Direct investigations of serum testosterone levels and genetic susceptibility to high testosterone production or sensitivity in women with ASC would illuminate the origin of these conditions. The relationship between FT and current testosterone levels also needs to be clarified. The present results may be relevant to understanding the increased male risk to developing autism.

Tumor liberated protein (TLP): its potential for diagnosis and therapy.

Our earlier studies led to the purification of a similar antigen from several types of human tumours which had the following properties: It provoked a delayed reaction in certain cancer patients which could be demonstrated both in vivo and in vitro (1). In contrast, healthy individuals did not react to this antigen. Subsequent studies led to a higher degree of purity of tumour antigen which enabled a partial sequence (2). The eight amino acid sequence obtained led to the synthesis of the corresponding peptide. This synthetic peptide was then used to raise specific antibodies in rabbits. The availability of such antibodies enabled to undertake a range of studies. Those investigations revealed that the antibodies were tumour specific because they reacted only with certain types of tumours but not with normal cells nor with other types of tumour tissues. We decided to name this tumour antigen as TLP for tumour liberated protein.

[Detection of urogenital malignant cells in the peripheral blood by nested RT-PCR using keratin 19 mRNA].

INTRODUCTION AND OBJECTIVES: It is reported that cytokeratin 19 (CK-19) mRNA is not expressed in the peripheral blood cells of the healthy subjects (Am. J. Pathol. 142: 1111. 1993). Detection of DNA fragments of CK-19 in the peripheral blood suggests the existence of epithelial malignant tumor cells. In this study, we detected CK-19 genes in peripheral blood of patients with urogenital malignancy, and thus clarified the possibility of understanding tumor expansion. METHODS: Mononuclear cells were separated from the peripheral blood of 39 patients with urogenital tumor and 9 controls. Total cellular RNA was extracted according to the method described elsewhere. The CK-19 gene expression was investigated using nested reverse transcription polymerase chain reaction (nested RT-PCR) and confirmed by Southern Blotting. These results were compared to the clinical stages. RESULTS: CK-19 mRNA was not detected in any of 9 controls. CK-19 mRNA was detected in 3 out of the 10 bladder cancer cases. Two had metastases and 1 had muscle invasion without metastasis. One of 6 urothelial cancer case of the upper urinary tract showed positive amplification, and had liver, bone and lymphnode metastases. After one course of chemotherapy, mRNA became negative. In prostate cancer cases, three among 7 were positive and all 3 cases had distant metastases. In renal, testicular and penile cancer cases, the positive ratio were 3/6, 1/8 and 2/2, respectively. All positive cases of renal and testicular cancer had distant metastases, but one case of penile cancer had no metastasis. CONCLUSION: The detection of CK-19 mRNA in the circulating blood by nested RT-PCR makes it possible to detect micrometastasis, to evaluate therapeutic effects and to predict the prognosis.

Studies on tumourous and other urogenital patients with respect to antigens of oncogenic adenovirus.

The possible connection of viruses with tumours was investigated by serologic examinations. Concerning the presence of antibodies against adenoviruses, especially those against the early non-virion antigens of oncogenic adenovirus type 12, approximately 4000 tests were made with sera of 446 urogenital patients with and without tumours and 70 ones with internal diseases. It was found by complement fixation tests that antibodies against nonvirion antigens of adenoviruses were present in 53% of urogenital patients suffering from malignant tumours and prostatic hypertrophy, in 18% of non-tumourous urological patients and in 4% of patients with internal diseases, respectively. The results suggest that adenoviruses may play a role in tumourous diseases of the urogenital organs.

[Bone metastasis in patients with urogenital neoplasms]. / Przerzuty do kosci u chorych na nowotwory narzadów ukladu moczowo-plciowego.

The clinical data of 91 patients with bone metastases were reviewed. The renal cell carcinoma and prostatic carcinoma were diagnosed respectively in 53% and 47% of the patients. 48% of the patients had tumour size stage T3 and 71% had histopathological stage II (G2). 21% of the patients presented a bone pain. In patients with renal cell carcinoma, the level of serum bone alkaline phosphatase and erythrocyte sedimentation rate were correlated with the concentration of serum ferritin (respectively p = 0.008 and p = 0.055). The relationship between the histopathological grade (G) and the stage of tumour size (T), and the concentration of serum ferritin was noted. In patients with prostatic carcinoma, the relationship between general condition and the concentration of prostatic specific antigen (PSA) as well as the relationship between PSA and the intensity of bone pain were observed. Only relationship between the histopathological grade and the concentration of PSA had a statistical significance (p < 0.05).

Determination of serum total lipid and free N-acetylneuraminic acid in genitourinary malignancies by fluorimetric high performance liquid chromatography. Relevance of free N-acetylneuraminic acid as tumour marker.

The reliability of total sialic acid (TSA), lipid sialic acid (LSA) and free sialic acid (FSA) as markers in genitourinary malignancies was evaluated in 20 normal subjects, 21 patients with prostatic cancer, 22 patients with urinary bladder cancer and 14 patients with renal cell carcinoma. We introduce the new concept of 'corrected' lipid sialic acid (CLSA), which expresses the actual concentration of sialic acid bound to glycolipds by subtracting the concentration of FSA determined by a novel ultrafiltration method. TSA did not show significant differences with respect to normal controls, except for renal cell carcinoma, whose mean value (879 +/- 145 micrograms/ml) showed a P value < 0.001. Instead, CLSA showed only significant differences (P = 0.001), with respect to normal controls in stage I and in all grades of renal cell carcinoma. While all data indicated significant increases (P = 0.001) in the FSA values, (means +/- S.D.) of 0.621 +/- 0.272 micrograms/ml were found in patients with prostatic cancer, 0.796 +/- 0.443 micrograms/ml in patients with urinary bladder cancer and 0.667 +/- 0.146 micrograms/ml in patients with renal cell carcinoma. Separate TSA and CLSA measurements appeared to be of limited value in the detection of genitourinary malignancies. However, results show that FSA was the most sensitive of the three markers tested for detecting malignancies.