Integrative immunologic and genomic characterization of brain metastasis from ovarian/peritoneal cancer

Brain metastasis from ovarian/peritoneal cancer is a rare disease that has a dismal prognosis. And genomic alterations and immune-profiling in primary ovarian/ peritoneal cancer and brain metastatic tumor tissues have not been fully elucidated. Multiplexed immunofluorescence and whole-exome sequencing of two matched brain metastatic tumor and primary ovarian/peritoneal cancer tissues were performed. The overall density of immune infiltrates in metastatic tissues (brain) was not significantly different from those in primary cancer tissues (case 1 primary: 2.12% and case 1 metastasis: 2.22%; case 2 primary: 1.70%, and case 2 metastasis: 3.46%). Of note, however, PD-L1 expression in the metastases was higher than that in the primary tumors. We found more non-silent mutations, cancer-related genes, loss of heterozygosity (LOH) and longer lengths of copy-number alterations (CNA) in brain metastases compared to primary ovarian/peritoneal cancers. We report immunologic and genomic profiles of primary ovarian/peritoneal cancer with brain metastasis that may not only provide useful information for understanding its pathogenesis, but also clues for further innovative therapeutic treatments for ovarian cancer.     

Characteristic location and rapid progression of medial femoral condylar chondral lesions accompanying medial meniscus posterior root tear

BackgroundThis study aimed to demonstrate the characteristics of the chondral lesion at the medial femoral condyle (MFC) in patients with medial meniscal posterior root tear (MMPRT), and to compare the progression rate of accompanying chondral disease in MMPRT with that in degenerative non-radial tear.MethodsThirty-one patients who underwent arthroscopic surgery for MMPRT and 31 controls who underwent arthroscopic surgery for degenerative medial meniscus posterior horn horizontal tear (MMPHT) were included. Accompanying chondral lesions in the MFC were evaluated by magnetic resonance imaging (MRI) at initial diagnosis and from video taken at arthroscopic surgery using the International Cartilage Repair Society (ICRS) classification system. The difference in severity and extent of the chondral lesion between initial diagnosis and surgery was measured.ResultsTwenty-five patients with MMPRT (80.6%) and 29 patients (93.5%) with MMPHT had ICRS grade ≥ 2 chondral injuries at MFC. In the MMPRT and MMPHT groups, the most severely injured areas of the MFC were the far medial compartment (52%) and central compartment (51%), respectively. Comparing MRI and arthroscopy, 12 patients (39%) in the MMPRT group showed progression of chondral disease after a mean of 3.5 months, whereas only three patients (10%) in the MMPHT group showed progression after a mean of 3.1 months.ConclusionsMFC lesions accompanying MMPRT were located more medially and progressed faster than those with non-root horizontal tear. Earlier intervention such as repair of MMPRT or high tibial osteotomy, if malalignment is present, may be considered in order to prevent the progression of chondral lesion.     

BRAF and NRAS mutations and antitumor immunity in Korean malignant melanomas and their prognostic relevance: Gene set enrichment analysis and CIBERSORT analysis

Cutaneous malignant melanomas (CMMs) are rare but are the cause of the highest skin cancer-related mortality in Korea. Very few studies have investigated the associations between KRAS, NRAS, BRAF, and PIK3CA mutations and TICs, as well as their prognostic impact on Korean CMMs. Peptide nucleic acid-mediated polymerase chain reaction clamping and Mutyper and immunohistochemistry were used to detect these mutations in 47 paraffinized CMMs. BRAF and NRAS mutations were detected in 21.3% and 12.8% of CMMs, respectively. No KRAS or PIK3CA mutations were identified. NRAS mutations correlated with low FOXP3 (regulatory T lymphocyte marker) and indoleamine 2,3-dioxygenase (IDO) (activated dendritic cell marker) TICs in CMMs, which is consistent with the negative correlation of regulatory T cells with NRAS mutations in TCGA data, while BRAF mutations were not associated with any TICs. In gene set enrichment analysis, BRAF and NRAS mutations were enriched in decreased CD8 (suppressor/cytotoxic T lymphocyte marker) T cell-linked and increased CD4 (helper/inducer T lymphocyte marker) T cell-linked gene signatures, respectively, confirming the trend in our cohort of associations only with NRAS. BRAF or NRAS mutations alone did not affect any prognosis. In the subgroup analyses, BRAF mutations, as well as high CD4, CD8, FOXP3, and IDO TICs, caused worse overall survival in NRAS-mutated melanoma. No correlation of CD163 (monocyte–macrophage-specific marker) was detected.We found that approximately one-third of our cohort had BRAF and NRAS mutations, none had KRAS or PIK3CA mutations, and most displayed decreased anti-tumor immunity. These findings may warrant further study on combined immunotherapeutic and molecular targeted therapy in Korean CMMs. Subgroup analyses according to TICs and BRAF/NRAS mutations may help to identify high-risk patients with worse prognoses.     

Clinicopathological significance and prognostic implication of nuclear factor-κB activation in colorectal cancer

ObjectiveThe aim of the present study was to evaluate the clinicopathological significance of phosphorylated nuclear factor-κB (pNF-κB) expression, and its impact on epithelial–mesenchymal transition and angiogenesis in colorectal cancer (CRC).MethodsWe carried out immunohistochemistry of pNF-κB on 261 human CRC tissues, and evaluated nuclear expression, regardless of cytoplasmic expression. We also investigated the correlation between pNF-κB expression and clinicopathological characteristics, survival, and epithelial–mesenchymal transition and angiogenesis-related markers in CRC.ResultspNF-κB was expressed in the nuclei of 164 of the 261 CRC tissues (62.8%). Furthermore, pNF-κB was significantly correlated with frequent perineural invasion, lymph node metastasis, and higher pTNM stage. However, there was no significant correlation between pNF-κB expression and other clinicopathological parameters. Among the epithelial–mesenchymal transition markers examined, SNAIL expression was significantly correlated with pNF-κB expression (P = 0.001) but E-cadherin expression was not. CRC with pNF-κB expression had significantly higher SIRT1 expression levels and hypoxia-inducible factor-1α expression levels than CRC without pNF-κB expression (P < 0.001 and P < 0.001, respectively). However, there was no correlation between the expression levels of pNF-κB and VEGF. pNF-κB expression was significantly correlated with worse overall and recurrence-free survival rates (P < 0.001 and P < 0.001, respectively).ConclusionpNF-κB expression was significantly correlated with aggressive tumor behaviors and worse survival rates. Furthermore, pNF-κB expression may affect tumor invasion and progression through SNAIL-related epithelial–mesenchymal transition and SIRT1- and hypoxia-inducible factor-1α-induced angiogenesis.     

Size distribution of serum extracellular vesicles in mice with atherosclerosis

Background and aimsAtherosclerosis is a prominent vascular lesion, and potentially causing ischemic alterations in the brain and heart. Recent studies have reported that physiological and pathological alterations in atherosclerosis and extracellular vesicles (EV) are related. This study aimed to investigate the association between the extent of atherosclerotic lesions and the number of serum EVs in a mouse model of atherosclerosis (wild-type).MethodsEighteen 3-week-old C57BL/6 N male mice(wild-type) were purchased. Twelve mice were fed a 45% high-fat diet (HFD) for six months. Six mice were provided standard laboratory chow for six months. The entire aorta, from the aortic sinus to the division of the iliac artery, was dissected out from each mouse. Furthermore, the degree of atherosclerosis was microscopically determined. Serum EVs were quantified by size via nanoparticle tracking analysis.ResultsThe number of EVs in the high-atherosclerotic score group (1.43 × 10⁹) was higher than that in the low- atherosclerotic score group (0.7 × 10⁹) in the range of 211.5–222.5 nm (p = 0.033).ConclusionsEnumeration of EVs is a potential method of detecting atherosclerosis.     

Evolution of Klebsiella pneumoniae with mucoid and non-mucoid type colonies within a single patient

We obtained nine Klebsiella pneumoniae isolates successively isolated from a single patient. Four pairs (M1–M4 and NM1–NM4) obtained simultaneously from the same site showed different colony types, mucoid and non-mucoid, while the final isolate (M5) was isolated alone from the blood and showed a mucoid phenotype. The whole genome of isolate M5 was sequenced de novo using the PacBio RSII system, while the others were sequenced with an Illumina Hiseq4000 and mapped to the genome sequences of M5. To identify insertions or deletions in the cps locus, we amplified and sequenced cps locus genes. We identified insertion sequence (IS) elements in several genes of the cps locus or one amino acid substitution in WcaJ in all non-mucoid isolates. Five additional amino acid alterations in RpsJ, LolE, Lon-2, PpsE, and a hypothetical protein were detected in some mucoid and non-mucoid isolates. Based on the genome data and cps locus sequences, the mucoid phenotype may have been lost or converted into the non-mucoid phenotype because of the insertion of IS elements or amino acid alterations at this locus. We inferred a within-host evolutionary scenario, in which non-mucoid variants emerged repeatedly from mucoid isolates, but may be short-lived because of their low fitness.     

Gastric adenocarcinoma with enteroblastic differentiation should be differentiated from hepatoid adenocarcinoma: A study with emphasis on clear cells and clinicopathologic spectrum

BackgroundIn gastric cancer, clear cells are preferentially found in gastric adenocarcinoma with enteroblastic differentiation (GAED) and hepatoid adenocarcinoma (HAC). The distinction between GAED and HAC is difficult because of their rarity and histologic overlap.MethodsTo elucidate identification of gastric adenocarcinoma with clear cells as GAED or HAC, survival analyses were performed in 28 GAED, 26 HAC, and 1107 conventional adenocarcinoma cases. Cells of origin were assessed by investigating the expression of oncofetal proteins (α-FP, glypican-3, SALL4), in addition to gastric (MUC5AC, MUC6), and intestinal (MUC2, CD10, CDX-2) cell markers.ResultsClinically, HAC showed frequent (57.5%) distant metastasis (mostly in the liver) at the time of diagnosis compared to GAED (P < 0.001). On pathology, all 28 GAED had a predominantly tubulopapillary growth pattern while 24 HAC displayed a predominantly hepatoid growth pattern. In survival analyses, patients with HAC had significantly shorter overall and recurrence-free survival (mean: 25 months, and 53 months, respectively) compared to those with GAED (mean: 107 months, and 118 months, respectively) (P < 0.001).HAC with clear cells showed diffuse and strong expression of all oncofetal proteins (α-FP, glypican-3, and SALL4), were highly positive for CDX-2, and were negative for CD10, MUC6, MUC5AC, and MUC2, suggesting an intestinal mucin phenotype and hepatoid features. In contrast, GAED showed focal expression of one or two oncofetal proteins and commonly expressed CD10, CDX-2, and MUC6 but not MUC2 and MUC5AC, suggesting both gastric antral/intestinal mucin phenotype and focal enteroblastic differentiation. SALL4 was diffusely and strongly positive in HAC, while it was heterogeneously expressed in GAED.ConclusionsIn conclusion, although rare, HAC with clear cells should be differentiated from GAED based on the poor prognosis, diffuse and strong oncofetal protein expression, and intestinal mucin phenotype.