Histopathological correlation of punctate echogenic foci on ultrasonography in papillary thyroid carcinoma.

PURPOSE: To investigate the pathological entities of punctate echogenic foci (PEF) by correlating PEF with histopathological features of papillary thyroid carcinoma (PTC). METHODS: This study included 121 consecutive patients who had undergone thyroidectomy for PTC. The inclusion criterion was entire tumor resection with a 3-mm thickness for histopathological examination. We assessed the presence and number (<5 or ≥5) of PEF defined as punctate hyperechoic foci within the solid component of nodules. All surgical tumor specimens were retrospectively reviewed for the presence of microcalcifications, including the psammomatous calcification, coarse microcalcification, and micro-ossification, and inspissated colloid. RESULTS: PEF were detected in 71 (58.7%) PTCs. Psammomatous calcifications, coarse microcalcifications, and inspissated colloids were more frequently found in PTCs with PEF than in those without (74.6%, 42.3%, and 46.5%, respectively, p ≤ 0.024). Any type of microcalcification was found in 90.1% of PTCs with PEF. Psammomatous calcifications, coarse microcalcifications, and inspissated colloids were independently associated with PEF (p ≤ 0.012). Psammomatous calcifications were found in all PTCs with a high number (≥5) of PEF. CONCLUSION: Microcalcifications were found in most PTCs with PEF and psammomatous calcification was the main pathological entity of PEF in PTC. Our study validates reliability of PEF as a predictor of microcalcifications in PTC.

Immune classification for the PD-L1 expression and tumour-infiltrating lymphocytes in colorectal adenocarcinoma.

BACKGROUND: Colorectal adenocarcinoma is the third most common cancer worldwide and a leading cause of cancer-related death. The recent emergence of diverse immunotherapeutic agents has made it crucial to interpret a complex tumour microenvironment intermingled with tumour-infiltrating immune cells to predict the immunotherapeutic response rate. However, in colorectal adenocarcinoma, studies are lacking that provide detailed analyses of programmed death-ligand 1 (PD-L1) and tumour-infiltrating lymphocytes (TIL) to elucidate their prognostic values and to identify immunotherapy-targetable subgroups, preferably with multiple immune-related biomarkers. In the present study, we categorize colorectal adenocarcinomas into four types of tumour immune microenvironments according to PD-L1 expression and TIL, analyse their prognostic values, and propose an immunotherapy-targetable subgroup. METHODS: Formalin-fixed, paraffin-embedded tissue samples of surgically resected primary colorectal adenocarcinomas (n = 489) were obtained and arrayed on tissue microarray blocks. Immunohistochemical stains for PD-L1, programmed cell death protein 1 (PD-1), cluster of differentiation 8 (CD8), and deficient mismatch repair (dMMR) were performed and evaluated. RESULTS: Tumour microenvironment immune type (TMIT) I (PD-L1-positive tumour cells and CD8-high TIL) and type II (PD-L1-negative tumour cells and CD8-low TIL) showed the best and worst prognoses, respectively. PD-L1 overexpression was significantly associated with dMMR status. PD-L1 immunoreactivity was positively correlated with TIL having CD8 or PD-1 overexpression. CONCLUSIONS: TMIT I subgroup showed stronger CD8/PD-L1/PD-1 signalling interaction compared to the other TMIT. Therefore, we propose that the TMIT I subgroup is a candidate TMIT to predict effective response rate for existing immune checkpoint inhibitors and determine targetable subgroups for emerging therapies.

Preoperative ultrasonography parathyroid gland mapping can improve identification of normal parathyroid gland during thyroidectomy: A propensity score-matched case-control study.

BACKGROUND: Accurate intraoperative identification of normal parathyroid glands (PTGs) is vital to avoid hypocalcemia post total thyroidectomy. Although ultrasonography (US) has been shown to identify normal PTGs, the significance of preoperative US PTG mapping in this context is not well studied. This study evaluated the impact of preoperative US PTG mapping on intraoperative identification of normal PTGs during total thyroidectomy. METHODS: The study involved 161 consecutive patients who underwent total thyroidectomy between January 2020 and June 2022. These included patients without preoperative US PTG mapping (group 1, n = 91) and those with the mapping (group 2, n = 70). Propensity score matching yielded 61 matched patients from each group. We developed a preoperative US PTG mapping technique combining US identification of normal PTGs with their localization on thyroid CT images. The intraoperative detectability of normal PTGs during thyroid surgery and detectability of normal PTGs by the preoperative US mapping were assessed by the number of PTGs identified per patient and by location. RESULTS: In the matched cohort, group 2 demonstrated a higher median number of identified PTGs (3 vs. 2, p = 0.011), a greater proportion of patients with three or more identified PTGs (65.5% vs. 44.3%, p = 0.018), and a higher ratio of identified to expected PTGs (70.5% vs. 60.2%, p = 0.011) than group 1. In group 2, the median number of normal PTGs identified preoperatively was 3, with at least one identified in 95.7% of patients, two or more in 84.3%, three or more in 52.9%, and four or five in 24.3%. CONCLUSIONS: Preoperative US PTG mapping identified two or more normal PTGs in the majority of adult patients undergoing total thyroidectomy. Those with preoperative mapping showed a higher number of intraoperatively identified normal PTGs, including inferior PTGs, compared to those without. This technique appears to enhance the intraoperative identification of normal PTGs, thereby potentially improving surgical outcomes in total thyroidectomy.

US features of normal parathyroid glands: a comparison with metastatic lymph nodes in thyroid cancer.

PURPOSE: This study aimed to determine the ultrasound (US) features of normal parathyroid glands (PTGs) and to evaluate whether normal PTGs can be differentiated from metastatic lymph nodes (LNs) in thyroid cancer. METHODS: This retrospective study included 10 normal PTGs and 95 metastatic LNs from thyroid cancer showing suspicious US features. The echogenicity, echotexture, echogenic foci (calcifications), cystic change, abnormal vascularity, size, shape, and location were retrospectively assessed and compared between normal PTGs and metastatic LNs. RESULTS: The echogenicity of normal PTGs was significantly different from that of metastatic LNs (P<0.001). Normal PTGs exhibited marked hyperechogenicity (100%), homogeneous echotexture (80%), focal intraglandular hypoechogenicity (20%), ovoid shape (90%), and focal cystic change in one case (10%). The echogenicity of metastatic LNs was markedly hyperechoic (0%), moderately hyperechoic (15.8%), mildly hyperechoic (53.7%), and hypoechoic (28.4%). The size and long axis/short axis ratios of normal PTGs were significantly smaller and larger than those of metastatic LNs (P<0.01 and P=0.022, respectively). CONCLUSION: Marked hyperechogenicity was found only in normal PTGs, and small, ovoid, markedly hyperechoic structures in the paramedian central neck characterized normal PTGs. Normal PTGs may be differentiated from metastatic LNs in thyroid cancer.

US Features of the Parathyroid Glands: An Intraoperative Surgical Specimen Study.

Purpose: This study aimed to evaluate the US features of the parathyroid glands (PTGs) using surgical specimens of normal PTGs obtained during thyroid surgery. Materials and Methods: This study included 34 normal PTGs from 17 consecutive patients who underwent thyroid surgery between December 2020 and March 2021. All normal PTGs were histologically confirmed by intraoperative frozen-section biopsy for autotransplantation. Surgically resected parathyroid specimens were scanned in sterile normal saline using high-resolution US prior to autotransplantation. The US features of echogenicity (hyperechogenicity or hypoechogenicity), echotexture (homogeneous or heterogeneous), size, and shape (ovoid or round) were retrospectively evaluated. The echogenicity of the three PTGs was compared with that of the thyroid parenchyma of the resected thyroid specimens in two patients. Results: All PTGs showed hyperechogenicity similar to that of gauze soaked in normal saline. Homogeneous hyperechogenicity was observed in 32/34 (94.1%) patients, and the echogenicity of the three PTGs was hyperechoic compared with that of the thyroid parenchyma. The long diameter of the PTGs ranged from 5.1 mm to 9.8 mm (mean, 7.1 mm) and the shape of the PTGs was ovoid in 33/34 (97.1%) patients. Conclusion: The echogenicity of normal PTG specimens was consistently hyperechoic, and the small ovoid homogeneously hyperechoic structure was a characteristic US feature of the PTGs.

Dome-type carcinoma of the rectum mimicking a submucosal tumor: a case report and literature review.

Dome-type carcinoma (DC) has been recognized as a rare variant of adenocarcinoma, which arises in gut-associated lymphoid tissue. It has a specific morphologic feature of a dome-like protrusion associated with lymphoid tissue. We report a case of a DC of the rectum in an asymptomatic 58-year-old male. A 2-cm sized, well-demarcated, round mass masquerading as a submucosal tumor (SMT) was identified in the rectum and was resected by endoscopic submucosal dissection. The tumor was revealed as an adenocarcinoma with submucosal invasion of 3,700 µm, which consisted of dilated cystic glands and the lymphoid stroma with reactive germinal centers. On immunohistochemistry, the tumor cells revealed retained expression for mismatch repair proteins. Laparoscopic surgical resection was subsequently performed. DC is considered a distinctive subtype of colorectal adenocarcinoma with characteristic morphology and low-grade malignant potential. Careful detection of the overlying mucosal lesion is crucial to differentially diagnose DC from SMT.

Prognostic implications of immune classification using IDO1 expression in extrahepatic bile duct carcinoma.

Indoleamine 2, 3-dioxygenase 1 (IDO1) is an immunomodulatory enzyme that catalyzes the degradation of tryptophan to kynurenine and induces immune tolerance in tumor cells. The effects of IDO1 on extrahepatic bile duct carcinoma (EHBDC) are poorly understood. Therefore, the present study aimed to investigate the expression and prognostic significance of IDO1 in EHBDC. An immunohistochemical microarray analysis of IDO1 expression was performed for 76 surgically resected cases of EHBDC. CD8+ tumor infiltrating lymphocytes (TILs) were also investigated through a combination analysis with IDO1 expression. IDO1 was highly expressed in 25 of 76 (32.9%) cases. High expression of IDO1 was associated with decreased numbers of CD8+ TILs (P=0.008), a higher pN category (P=0.007), an advanced overall stage (P=0.001) and frequent recurrence (P=0.018). When IDO1 expression was further stratified with CD8+ TIL state, the IDO1high/CD8low subgroup was decreased in terms of overall survival (P=0.025) and disease-free survival (P=0.015) compared with IDO1high/CD8high, IDO1low/CD8high and IDO1low/CD8low subgroups. High IDO1 expression was associated with a decreased number of CD8+ TILs and associated with a poor prognosis. As IDO1 may be a new target of immunotherapy applications, IDO1/CD8+ TIL subgrouping can be a useful prognostic and predictive tool in patients with EHBDC.

Novel Biomarker Panels Discriminating between Pancreatic Ductal Adenocarcinoma and Extrahepatic Bile Duct Carcinoma.

OBJECTIVE: Pancreatic ductal adenocarcinoma (PDAC) and extrahepatic bile duct carcinoma (EBDC) are distinct entities with different clinicopathological implications. Therefore, research to differentiate between the two diseases is compulsory. In this study, four biomarkers were selected (Hippocalcin-like 1 (HPCAL1); annexin A10 (ANXA10); MUC5AC; sodium/potassium-transporting ATPase subunit beta-1 (ATP1B1)) and focus was placed on clarifying the diagnostic performance of each biomarker and pioneering novel-combined biomarker panels to discriminate between PDAC and EBDC. PROCEDURES: An immunohistochemical microarray analysis of HPCAL1, ANXA10, MUC5AC, and ATP1B1 was conducted for surgically resected 55 PDACs and 77 EBDCs. The diagnostic performance discriminating between PDAC and EBDC was evaluated using four biomarkers and the combined biomarker panels. RESULTS: PDACs exhibited more positive expressions for HPCAL1, ANXA10, and MUC5AC, whereas EBDCs exhibited more ATP1B1-positive expressions. The PDAC panel with the best diagnostic performance was the profile of (+ in ≥ 2 among HPCAL1, ANXA10, MUC5AC)/ATP1B1-. The immunophenotype pattern of (- in ≥ 1 among HPCAL1, ANXA10, MUC5AC)/ATP1B1+ is the EBDC panel with the most excellent discriminating power. CONCLUSION: The suggested combined biomarker panels demonstrate the distinguishing diagnostic ability between PDAC and EBDC is better than previous studies. Therefore, for differentiation between PDAC and EBDC, these panels are expected to help unravel the clinicopathological enigma as promising biomarker panels in the future.

Identification of New Prognostic Markers and Therapeutic Targets for Non-Muscle Invasive Bladder Cancer: HER2 as a Potential Target Antigen.

Bacillus Calmette-Guérin (BCG) is the gold standard adjuvant treatment for non-muscle-invasive bladder cancer (NMIBC). However, given the current global shortage of BCG, new treatments are needed. We evaluated tumor microenvironment markers as potential BCG alternatives for NMIBC treatment. Programmed death-ligand 1, human epidermal growth factor receptor-2 (HER2), programmed cell death-1 (PD1), CD8, and Ki67 levels were measured in treatment-naïve NMIBC and MIBC patients (pTa, pT1, and pT2 stages). Univariate and multivariate Cox proportional hazard models were used to determine the impact of these markers and other clinicopathological factors on survival, recurrence, and progression. EP263, IM142, PD1, and Ki67 levels were the highest in the T2 stage, followed by the T1 and Ta stages. HER2 and IM263 expressions were higher in the T1 and T2 stages than in the Ta stage. In NMIBC, the significant prognostic factors for recurrence-free survival were adjuvant therapy, tumor grade, and HER2 positivity, whereas those for progression-free survival included age, T-stage, and IM263. Age, T-stage, EP263, PD1, CD8, and Ki67 levels were significant factors associated with overall survival. IM263 and HER2 are potential biomarkers for progression and recurrence, respectively. Therefore, we propose HER2 as a potential target antigen for intravesical therapeutics as a BCG alternative.

Malignancy risk of thyroid nodules with nonshadowing echogenic foci.

PURPOSE: This study was conducted to determine the malignancy risk and diagnostic value of various types of nonshadowing echogenic foci (NEF) in the risk stratification of thyroid nodules. METHODS: A total of 1,018 consecutive thyroid nodules (≥1 cm) with final diagnoses were included. The presence of NEF was determined and types of NEF were classified according to the presence of a comet tail artifact (CTA), location, and size through a prospective evaluation. The associations with malignancy, malignancy risk, and diagnostic value of various types of NEF were assessed. RESULTS: Intrasolid punctate NEF without CTA was the only type of NEF that was an independent predictor of malignancy (P<0.001). The malignancy risk of intrasolid punctate NEF without CTA was substantially higher in solid hypoechoic nodules than in isoechoic or nonsolid nodules (71.3% vs. 9.2%, P<0.001). In solid hypoechoic nodules, slightly increased sensitivity (70.8% vs. 67.9%) for malignancy and a similar malignancy risk (71.4% vs. 71.3%) were observed for intrasolid punctate NEF (with or without CTA) and intrasolid punctate NEF without CTA, respectively. NEF with CTA at the margin of the cystic component was not associated with malignancy or benignity in nonsolid nodules (P>0.05). CONCLUSION: Intrasolid punctate NEF without CTA was the only independent predictor of malignancy. However, solid hypoechoic nodules with intrasolid punctate NEF should be classified as high-suspicion nodules regardless of coexisting CTA. Other types of NEF had no added value for detecting malignancy compared to intrasolid punctate NEF without CTA.

Malignancy Risk Stratification of Thyroid Nodules with Macrocalcification and Rim Calcification Based on Ultrasound Patterns.

OBJECTIVE: To determine the association of macrocalcification and rim calcification with malignancy and to stratify the malignancy risk of thyroid nodules with macrocalcification and rim calcification based on ultrasound (US) patterns. MATERIALS AND METHODS: The study included a total of 3603 consecutive nodules (≥ 1 cm) with final diagnoses. The associations of macrocalcification and rim calcification with malignancy and malignancy risk of the nodules were assessed overall and in subgroups based on the US patterns of the nodules. The malignancy risk of the thyroid nodules was categorized as high (> 50%), intermediate (upper-intermediate: > 30%, ≤ 50%; lower-intermediate: > 10%, ≤ 30%), and low (≤ 10%). RESULTS: Macrocalcification was independently associated with malignancy in all nodules and solid hypoechoic (SH) nodules (p < 0.001). Rim calcification was not associated with malignancy in all nodules (p = 0.802); however, it was independently associated with malignancy in partially cystic or isoechoic and hyperechoic (PCIH) nodules (p = 0.010). The malignancy risks of nodules with macrocalcification were classified as upper-intermediate and high in SH nodules, and as low and lower-intermediate in PCIH nodules based on suspicious US features. The malignancy risks of nodules with rim calcification were stratified as low and lower-intermediate based on suspicious US features. CONCLUSION: Macrocalcification increased the malignancy risk in all and SH nodules with or without suspicious US features, with low to high malignancy risks depending on the US patterns. Rim calcification increased the malignancy risk in PCIH nodules, with low and lower-intermediate malignancy risks based on suspicious US features. However, the role of rim calcification in risk stratification of thyroid nodules remains uncertain.

Prognostic implications of immune classification in a multicentre cohort of patients with small intestinal adenocarcinoma.

The diagnosis of small intestinal adenocarcinoma (SIAC) is usually determined at an advanced stage due to non-specific symptoms and the difficulty of exploring the small intestine. Therefore, the majority of SIAC patients have limited chemotherapeutic options. Until recently, the development of novel and effective therapies for SIAC have been limited owing to the low number of samples that have been collected and the low incidence of SIAC. Immunotherapies are becoming a focus. However, in SIAC, only a few studies to identify immunotherapy-responsive subgroups and their prognostic indicators have been reported. In the present study, we categorise primary SIAC into four types of tumour immune microenvironments and propose a strategy for identifying patient subgroups that are most likely to be immunotherapy-responsive. Formalin-fixed, paraffin-embedded tissue samples of a multicentre cohort of patients with SIAC (n=195) were collected using tissue microarrays. Immunohistochemical (IHC) stains for PD-L1, PD-1, and CD8 were performed, and microsatellite instability was evaluated using an IHC stain. Tumour microenvironment immune type (TMIT) I [PD-L1-positive tumour cells and CD8-high tumour-infiltrating lymphocytes (TILs)] and TMIT III (PD-L1-positive tumour cells and CD8-low TILs) show the best and worse prognoses, respectively. PD-L1 expression was significantly associated with high microsatellite instability (MSI) status. CD8-high TILs were positively correlated with PD-1-high TILs and high MSI. The TMIT I subgroup demonstrated a more patent CD8/PD-L1/PD-1 signalling pathway compared to other TMITs. Therefore, the TMIT I subgroup can be expected to have an effective response to immune checkpoint inhibitor therapies in SIAC. Such classification of SIACs into four immune types can be useful in predicting the prognosis of patients and the identification of immunotherapy-responsive subgroups.

Thyroid Nodules with Isolated Macrocalcifications: Malignancy Risk of Isolated Macrocalcifications and Postoperative Risk Stratification of Malignant Tumors Manifesting as Isolated Macrocalcifications.

OBJECTIVE: To determine the malignancy risk of isolated macrocalcifications (a calcified nodule with complete posterior acoustic shadowing) detected on ultrasonography (US) and to evaluate the postoperative American Thyroid Association (ATA) risk stratification of malignant tumors manifesting as isolated macrocalcifications. MATERIALS AND METHODS: A total of 3852 thyroid nodules (≥ 1 cm) of 3061 consecutive patients who had undergone biopsy between January 2011 and June 2018 were included in this study. We assessed the prevalence, malignancy rate, and size distribution of isolated macrocalcifications and evaluated the histopathologic features and postoperative ATA risk stratification of malignant tumors manifesting as isolated macrocalcifications. RESULTS: Isolated macrocalcifications were found in 38 (1.2%) of the 3061 patients. Final diagnosis was established in 30 (78.9%) nodules; seven malignant tumors were diagnosed as papillary thyroid carcinomas (PTCs). The malignancy rate of the isolated macrocalcifications was 23.3% in the 30 nodules with final diagnoses and 18.4% in all nodules. Among the six surgically-treated malignant tumors, five (83.3%) had an extrathyroidal extension (ETE) (minor ETE 1, gross ETE 4), and two (33.3%) had macroscopic lymph node metastasis. Four (66.7%) malignant tumors were categorized as high-risk tumors, one as an intermediate-risk tumor, and one as a low-risk tumor using the ATA risk stratification. Histopathologically, out of the six malignant tumors, ossifications were noted in four (66.7%) and predominant calcifications in two (33.3%). CONCLUSION: The US pattern of isolated macrocalcifications (≥ 1 cm) showed an intermediate malignancy risk (at least 18.4%). All malignant tumors were PTCs, and most showed an aggressive behavior and a high or intermediate postoperative ATA risk.

Phosphorylation of TFCP2L1 by CDK1 is required for stem cell pluripotency and bladder carcinogenesis.

Molecular programs involved in embryogenesis are frequently upregulated in oncogenic dedifferentiation and metastasis. However, their precise roles and regulatory mechanisms remain elusive. Here, we showed that CDK1 phosphorylation of TFCP2L1, a pluripotency-associated transcription factor, orchestrated pluripotency and cell-cycling in embryonic stem cells (ESCs) and was aberrantly activated in aggressive bladder cancers (BCs). In murine ESCs, the protein interactome and transcription targets of Tfcp2l1 indicated its involvement in cell cycle regulation. Tfcp2l1 was phosphorylated at Thr177 by Cdk1, which affected ESC cell cycle progression, pluripotency, and differentiation. The CDK1-TFCP2L1 pathway was activated in human BC cells, stimulating their proliferation, self-renewal, and invasion. Lack of TFCP2L1 phosphorylation impaired the tumorigenic potency of BC cells in a xenograft model. In patients with BC, high co-expression of TFCP2L1 and CDK1 was associated with unfavorable clinical characteristics including tumor grade, lymphovascular and muscularis propria invasion, and distant metastasis and was an independent prognostic factor for cancer-specific survival. These findings demonstrate the molecular and clinical significance of CDK1-mediated TFCP2L1 phosphorylation in stem cell pluripotency and in the tumorigenic stemness features associated with BC progression.

Pathogenetic implications of early growth response 1 in Ewing sarcoma.

Ewing sarcoma (ES) is the second most common primary malignant bone tumour, mainly occurs in children and adolescents, and has an overwhelming mortality. Despite extensive studies, few effective oncogenic signals have been described. Therefore, it is crucial to exploit novel pathognomonic factors and targetable biomarkers for ES patients. Based on previous studies, we speculate that insulin-like growth factor 1 receptor (IGF1R), which is upregulated by early growth response 1 (EGR1), may play a pivotal role in strengthening the downward transmission of IGF1 cascades. Therefore, in this study, we concentrated on determining the pathogenetic contribution of EGR1 in diverse ES cells. This report is the first to study the pathogenic role of EGR1 in ES. ES cells were cultured and transfected with Stealth RNAi human EGR1 small interfering RNA (siRNA) or negative control. Cell proliferation and invasion potential were measured. mRNA and protein expression of EGR1, IGF1R, and EWS-FLI1 also were assessed. In all EGR1 siRNA-transfected cells (SK-ES-1, RD-ES, and HS863.T), cell proliferation and invasive potential decreased significantly in EGR1 siRNA-transfected ES cells. mRNA and protein expression for EGR1, IGF1R, and EWS-FLI1 were also significantly reduced. In conclusion, EGR1 upregulated IGF1R expression and enhanced the expression of the oncogenic fusion protein EWS-FLI1. The EWS-FLI1/EGR1/IGF1R cascade combined with the previously confirmed pathways can form a speculative circuit, implicating positive feedback for tumourigenesis in ES. Therefore, EGR1 inhibitors are expected to be useful for the treatment of ES by preventing oncogenic IGF1/IGF1R expression.

Giant cell tumor of bone: updated molecular pathogenesis and tumor biology.

Giant cell tumor of bone (GCTB)-related clonal aberrations occur in a background of epigenetic histone modifications (especially the G34W mutation of H3F3A gene) that induce cytogenetic abnormalities. Clonal aberrations are closely linked to the aggressiveness of GCTB. The "neoplastic" mononuclear stromal cells in GCTB express fundamental RANKLs and various chemokines and cytokines associated with monocyte recruitment and "reactive" multinucleated giant cells (osteoclastogenesis). The reciprocal and orchestrated actions between mononuclear stromal cells and multinucleated giant cells help in the understanding of the molecular pathogenesis and tumor biology of GCTB. In the future, novel targets in the updated tumor biology and molecular pathogenesis of GCTB should be explored and scrutinized for the development of systemic therapy.

Prognostic Significance of Categorizing Gastric Carcinoma by PD-L1 Expression and Tumor Infiltrating Lymphocytes.

GOALS: We categorized gastric cancers into four types of tumor immune microenvironment based on PD-L1 expression and tumor-infiltrating lymphocytes (TILs) to analyze clinicopathologic characteristics and recommend an immunotherapy-targetable subgroup. PROCEDURES: Formalin-fixed, paraffin-embedded tissue samples of randomly selected gastric adenocarcinoma (n=479) were obtained and arrayed using a tissue-arraying instrument. Immunohistochemical stains for PD-L1, PD-1, and CD8 were performed and evaluated in addition to microsatellite instability, EBV infection, and HER-2 analyses. RESULTS: Tumor microenvironment immune type (TMIT) I (both PD-L1-positive tumor cells & CD8-high TILs) and type III (PD-L1-positive tumor cells & CD8-low TILs) showed the best and worse prognosis, respectively. PD-L1 expression was significantly associated with Epstein-Barr virus infection (p<0.001) and microsatellite instability status (p<0.001). PD-L1 immunoreactivity was positively correlated with TILs having CD8 or PD-1 overexpression. CONCLUSIONS: TMIT I subgroup showed more patent CD8/PD-L1/PD-1 signaling pathway compared to other types. Therefore, TMIT I subgroup is a good candidate to predict effective response rate for immune checkpoint blockers. Such stratification of gastric cancers into four types can be used to predict the prognosis of patients and determine the immunotherapy-targetable subgroup.

CCAR2 negatively regulates IL-8 production in cervical cancer cells.

Cell cycle and apoptosis regulator 2 (CCAR2) is a multifaceted protein that controls diverse cellular functions; however, its function in cancer is unclear. To better understand its potential role in cancer, we examined gene expression patterns regulated by CCAR2 in cervical cancer cells. Cytokine and chemokine production by CCAR2-deficient cells increased under oxidative conditions. In particular, H2O2-treated CCAR2-depleted cells showed a significant increase in interleukin-8 (IL-8) production, indicating a negative regulation of IL-8 by CCAR2. Upregulation of IL-8 expression in CCAR2-deficient cells occurred via activation of transcription factor AP-1. The negative correlation between CCAR2 and IL-8 expression was confirmed by examining mRNA and protein levels in tissues from cervical cancer patients. Furthermore, CCAR2-regulated IL-8 expression is associated with a shorter survival of cervical cancer patients. Overall, the data suggest that CCAR2 plays a critical role in controlling both the cancer secretome and cancer progression.

Improved efficacy and in vivo cellular properties of human embryonic stem cell derivative in a preclinical model of bladder pain syndrome.

Interstitial cystitis/bladder pain syndrome (IC/BPS) is an intractable disease characterized by severe pelvic pain and urinary frequency. Mesenchymal stem cell (MSC) therapy is a promising approach to treat incurable IC/BPS. Here, we show greater therapeutic efficacy of human embryonic stem cell (hESC)-derived multipotent stem cells (M-MSCs) than adult bone-marrow (BM)-derived counterparts for treating IC/BPS and also monitor long-term safety and in vivo properties of transplanted M-MSCs in living animals. Controlled hESC differentiation and isolation procedures resulted in pure M-MSCs displaying typical MSC behavior. In a hydrochloric-acid instillation-induced IC/BPS animal model, a single local injection of M-MSCs ameliorated bladder symptoms of IC/BPS with superior efficacy compared to BM-derived MSCs in ameliorating bladder voiding function and histological injuries including urothelium denudation, mast-cell infiltration, tissue fibrosis, apoptosis, and visceral hypersensitivity. Little adverse outcomes such as abnormal growth, tumorigenesis, or immune-mediated transplant rejection were observed over 12-months post-injection. Intravital confocal fluorescence imaging tracked the persistence of the transplanted cells over 6-months in living animals. The infused M-MSCs differentiated into multiple cell types and gradually integrated into vascular-like structures. The present study provides the first evidence for improved therapeutic efficacy, long-term safety, and in vivo distribution and cellular properties of hESC derivatives in preclinical models of IC/BPS.