Search

Supplementary figure 1 with 4 panels of images

Insulin-like growth factor binding protein 3 (IGFBP3) modulates cell growth through IGF-dependent and -independent mechanisms. Reports suggest that the serum levels of IGFBP3 are associated with various cancers and that IGFBP3 expression is significantly decreased in cisplatin (CDDP)-resistant lung cancer cells. Based on these findings, we investigated whether Igfbp3 deficiency accelerates mouse lung tumorigenesis and if expression of IGFBP3 enhances CDDP response by focusing on the IGF1 signaling cascade. To this end, an Igfbp3-null mouse model was generated in combination with KrasG12D to compare the tumor burden. Then, IGF-dependent signaling was assessed after expressing wild-type or a mutant IGFBP3 without IGF binding capacity in non–small cell lung cancer (NSCLC) cells. Finally, the treatment response to CDDP chemotherapy was evaluated under conditions of IGFBP3 overexpression. Igfbp3-null mice had increased lung tumor burden (>2-fold) and only half of human lung cancer cells survived after expression of IGFBP3, which corresponded to increased cleaved caspase-3 (10-fold), inactivation of IGF1 and MAPK signaling. In addition, overexpression of IGFBP3 increased susceptibility to CDDP treatment in lung cancer cells. These results, for the first time, demonstrate that IGFBP3 mediates lung cancer progression in a KrasG12D mouse model. Furthermore, overexpression of IGFBP3 induced apoptosis and enhanced cisplatin response in vitro and confirmed that the suppression is in part by blocking IGF1 signaling.Implications: These findings reveal that IGFBP3 is effective in lung cancer cells with high IGF1 signaling activity and imply that relevant biomarkers are essential in selecting lung cancer patients for IGF1-targeted therapy. Mol Cancer Res; 15(7); 896–904. ©2017 AACR.

We hypothesized that pancreatic and periampullary adenocarcinoma recurrence after surgical resection may be affected by the shedding of malignant epithelial cells during surgical dissection and that this may have implications for disease recurrence and survival.In this ongoing, investigator-initiated prospective randomized controlled trial, patients with pancreatic and periampullary adenocarcinoma were randomized intraoperatively, postresection into 3 study arms: peritoneal lavage using 10 L normal saline or distilled water, or control group with no lavage. Peritoneal fluid was sampled for cytologic analysis (cytospin, cellblock, immunohistochemistry-Ber-EP4 antibody) at 4 stages: (1) abdominal entry pre-dissection, (2) resection bed after tumor extirpation, (3) ex vivo resected specimen, and (4) resection bed postlavage.Between April 2016 and May 2018, 193 patients who underwent randomization for the study also underwent the described cytologic sampling. Of these, 167 patients (86.5%) were ultimately found to have pancreatic or periampullary adenocarcinoma. Before dissection (1) on cytospin analysis, 4.9% were positive, which rose to 10.2% intraoperatively (2), 16.7% ex vivo (3), and decreased to 4.3% (4) after lavage. Lymph node metastasis, margin involvement, and perineural invasion did not correlate with locoregional recurrence (LR). Tumor cells in the ex vivo cytospin (3) correlated with LR (odds ratio 3.8 [95% CI 1.6 to 9.2], p = 0.005) and LR disease-free survival (p = 0.007). Cox regression analysis revealed ex vivo cytospin positivity to be strongly associated with poorer LR disease-free survival (hazard ratio 2.26 [95% CI 1.16 to 4.42], p = 0.017).Cytologic sampling from ex vivo specimen irrigation after surgical resection of pancreatic and periampullary adenocarcinoma may have implications for LR, survival, and treatment, suggesting a possible cancer cell shedding phenotype.

To analyze CD8+ and FoxP3+ T-cell cellular density (CD) and intercellular distances (ID) in head and neck squamous cell carcinoma (HNSCC) samples from a neoadjuvant trial of durvalumab +/- metformin.Paired pre- and post-treatment primary HNSCC tumor samples were stained for CD8+ and FoxP3+. Digital image analysis was used to determine estimated mean CD8+ and FoxP3+ CDs and CD8+-FoxP3+ IDs in the leading tumor edge (LTE) and tumor adjacent stroma (TAS) stratified by treatment arm, human papillomavirus (HPV) status, and pathologic treatment response. A subset of samples was characterized for T-cell related signatures using digital spatial genomic profiling.Post-treatment analysis revealed a significant decrease in FoxP3+ CD and an increase in CD8+ CDs in the TAS between patients receiving durvalumab and metformin versus durvlaumab alone. Both treatment arms demonstrated significant post-treatment increases in ID. Although HPV+ and HPV- had similar immune cell CDs in the tumor microenvironment, HPV+ pre-treatment samples had 1.60 times greater ID compared with HPV- samples, trending toward significance (p = 0.05). At baseline, pathologic responders demonstrated a 1.16-fold greater CD8+ CDs in the LTE (p = 0.045) and 2.28-fold greater ID (p = 0.001) than non-responders. Digital spatial profiling revealed upregulation of FoxP3+ and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) in the TAS (p = 0.006, p = 0.026) in samples from pathologic responders.Analysis of CD8+ and FoxP3+ detected population differences according to HPV status, pathologic response, and treatment. Greater CD8+-FoxP3+ ID was associated with pathologic response. CD8+ and FoxP3+ T-cell distributions may be predictive of response to immune checkpoint inhibition.gov (Identifier NCT03618654).Level 3 Laryngoscope, 2022.

Despite optimal surgical resection, meningiomas may recur, with increasing grade and the degree of resection being predictive of risk. We hypothesize that an increasing Ki67 correlates with a higher risk of recurrence of resected WHO grade I meningiomas.The study population consisted of patients with resected WHO grade 1 meningiomas in locations outside of the base of skull. Digitally scanned slides stained for Ki67 were analyzed using automatic image analysis software in a standardized fashion.Recurrence was observed in 53 (17.7%) of cases with a median follow up time of 25.8 months. Ki67 ranged from 0 to 30%. Median Ki67 was 5.1% for patients with recurrence and 3.5% for patients without recurrence. In unadjusted analyses, high Ki-67 (≥ 5 vs.5) vs. ≥ 5) was associated with over a twofold increased risk of recurrence (13.1% vs. 27% respectively; HR 2.1731; 95% CI [1.2534, 3.764]; p = 0.006). After Adjusting for patient or tumor characteristics, elevated Ki-67 remained significantly correlated with recurrence. Grade 4 Simpson resection was noted in 71 (23.7%) of patients and it was associated with a significantly increased risk of recurrence (HR 2.56; 95% CI [1.41, 4.6364]; p = 0.002).WHO grade 1 meningiomas exhibit a significant rate of recurrence following resection. While Ki-67 is not part of the WHO grading criteria of meningiomas, a value greater than 5% is an independent predictor for increased risk of local recurrence following surgical resection.

Alterations of cellular metabolism have been implicated in immune dysfunction in the tumor microenvironment (TME) of head and neck squamous cell carcinoma (HNSCC). Metformin has recently emerged as a candidate of interest for combination with immunotherapy in HNSCC. This study investigated the effect of metformin on immune cell infiltrates of HNSCC.Retrospective analysis of T cell infiltrates in primary tumor specimens from patients enrolled in a clinical window of opportunity trial of presurgical metformin. Metformin was titrated to a standard diabetic dose (2000 mg/day) for a minimum of 9 days (mean 13.6 days) prior to surgical resection. Pre and posttreatment surgical specimens from 36 patients (16 HPVMetformin treatment was associated with a 41.4% decrease in FOXP3Metformin treatment may favorably alter the immune TME in HNSCC independent of HPV status.1b. This study is most accurately described as a non-randomized controlled trial and therefore may reflect a level of evidence below 1b but above 2a from the provided "levels of evidence" chart. Laryngoscope, 130:E490-E498, 2020.

Several lines of evidence strongly suggest that AAA is a specific antigen-driven T-cell inflammatory disease with a strong genetic component. We tested the hypothesis that γ- and δ-chain TCR transcripts are clonally expanded in AAA lesions. Previously, we showed clonally expanded α-chain (PloS One 2019;14:e0218990) and β-chain (J Immunol. 2014; 192:4897) TCR transcripts in AAA lesions of patients with AAA. We employed two-sided VγI-, VγII-, Vδ1- and Vδ2-specific PCR followed by cloning and sequencing of amplified transcripts to examine whether VγI-, VγII-, Vδ1- and Vδ2-TCR transcripts, respectively, were clonally expanded in aneurysmal lesions of patients with AAA. Clonally expanded VγI-, VγII-, Vδ1- and Vδ2-TCR transcripts were identified in AAA aneurysmal lesions. We identified multiple identical copies of: (i) VγI-TCR transcripts in 16 of 18 patients (range of identical transcripts 30% to 80% in individual patients); (ii) VγII-TCR transcripts in 15 of 15 patients (range of identical transcripts 39% to 80%);.(iii) Vδ1-TCR transcripts in 12 of 12 patients (range of identical transcripts 29% to 94%); and (iv) Vδ2-TCR transcripts in 10 of 10 patients (range of identical transcripts 40% to 91%). These results were statistically significant. PBMC from normal donors (methodological controls) contained almost exclusively unique VγI-, VγII- and Vδ2-TCR transcripts. In contrast, Vδ1 transcripts from PBMC from certain normal donors contained clonally expanded T cells, in agreement with previous reports. In conclusion, clonally expanded γδ TCR-expressing T cells were found in aneurysmal lesions of patients with AAA. These T cells may play an important role in the pathogenesis of AAA. Supported in part by grant RO1 HL64340 from NIH.

AAA is often a life-threatening disease with poorly understood pathogenesis. We examined gene expression profiles between AAA and normal aorta specimens by cDNA microarray analysis. AAA wall specimens from 5 patients with AAA and grossly normal aortic walls from 3 transplant donors were employed in this study. Total RNA was extracted from each specimen, RT into cDNA, and labeled with phosphorus α-32P. cDNA probes were then hybridized onto human cardiovascular microarray and analyzed by densitometry. Quantitative differences of some genes were further explored by SYBR green-mediated qRT-PCR analysis. Pairwise comparisons of both microarray and qRT-PCR results were made by Student t-test. Most of the 588 genes studied had similar expression among AAA specimens and control aortas, however, 25 genes were upregulated and 26 downregulated. Expression of MMP-9, TIMP3, collagen 1A1 (COL1A1), COL6A3, COL15A1, decorin, MCP 1, Apo E, Apo D, H19 and others was increased. Expression of COL4A4, biglycan, caveolin 3, galectin 1, LDL receptor-related protein 1, extracellular SOD 3, ANP receptor A precursor and others was decreased. Most qRT-PCR results agreed well with those from microarray analysis, with few exceptions. A number of genes that have been previously postulated to participate in AAA pathogenesis, have been upregulated or their expression has been altered. We also revealed some genes that have not been yet studied in human AAA. Of note, H19, one of the most highly up-regulated transcripts in two mouse aneurysm models, was also significantly upregulated in human AAA. These results indicate that many genes involved in different cellular functions might contribute to the pathogenesis of AAA. Supported in part by grant RO1 HL64340 from NIH.

This article covers, in a comprehensive way, thyroid cytopathology. The Bethesda System for Reporting Thyroid Cytology is reviewed, with emphasis on the atypical and indeterminate diagnostic categories. Immunohistochemistry stains and molecular tests panels applicable to cytology specimens are described.

Cancer patients undergo routine clinical monitoring with an array of blood tests that may carry long-term prognostic information. We aimed to develop a new prognostic model predicting survival for patients with advanced non-small cell lung cancer (NSCLC), based on laboratory tests commonly performed in clinical practice. A cohort of 1,161 stage IIIB or IV NSCLC patients was divided into training (n = 773) and testing (n = 388) cohorts. We analyzed the associations of 32 commonly tested laboratory variables with patient survival in the training cohort. We developed a model based on those significant laboratory variables, together with important clinical variables. The model was then evaluated in the testing cohort. Five variables, including albumin, total protein, alkaline phosphatase, blood urea nitrogen and international normalized ratio, were significantly associated with patient survival after stepwise selection. A model incorporating these variables classified patients into low-, medium- and high-risk groups with median survival of 16.9, 7.2 and 2.1 months, respectively (p0.0001). Compared with low-risk group, patients in the medium- and high-risk groups had a significantly higher risk of death at 1 year, with hazard ratio (HR) of 1.95 (95% CI 1.62-2.36) and 5.22 (4.30-6.34), respectively. These results were validated in the testing cohort. Overall, we developed a prognostic model relying entirely on readily available variables, with similar predictive power to those which depend on more specialized and expensive molecular assays. Further study is necessary to validate and further refine this model, and compare its performance to models based on more specialized and expensive testing.