Cases Of Colonic Adenocarcinoma Research Articles

Adenosquamous Carcinoma of the Proximal Colon: Case Report and Literature Review of a Rare, Aggressive Entity

Abstract Primary adenosquamous carcinoma of the colon is exceedingly rare and accounts for a clinically aggressive minority of colonic epithelial malignancies, with metastatic disease identified in 36% to 49% of cases at the time of diagnosis or resection, compared to 14% of conventional adenocarcinoma cases. Reported median overall survival time is less than half of that for conventional colonic adenocarcinoma, supporting inclusion of squamous differentiation as an adverse prognostic factor. Due to its rarity, optimal treatment protocols have yet to be standardized. We present the case of a 61-year-old female who underwent emergent right hemicolectomy due to bowel obstruction secondary to an ileocecal mass discovered on imaging. Gross examination revealed a 7.3-cm, primarily submucosal cecal mass involving the appendix with associated ileocolic adhesion. Histologic examination showed carcinoma with distinct areas of glandular and squamous differentiation, exhibiting the anticipated differential expression patterns of CK5/6, p63, CK20, CDX2, and mucicarmine. Immunohistochemistry for p16 was negative in both components. The adenocarcinoma component was continuous with a small focus of dysplastic colonic surface epithelium, supportive of cecal origin. Eight of 19 lymph nodes were positive for squamous and adenocarcinomatous metastasis, and a concurrent liver biopsy was also positive for metastasis. There was no evidence of mismatch repair protein deficiency by immunohistochemistry, which was performed on tissue containing both glandular and squamous components. We share this case to highlight a rare and uniquely aggressive subtype of a routinely encountered malignancy, reinforcing the importance of identifying and reporting squamous differentiation within cases of colonic adenocarcinoma.

ALPHLARD-NT: Bayesian Method for Human Leukocyte Antigen Genotyping and Mutation Calling through Simultaneous Analysis of Normal and Tumor Whole-Genome Sequence Data.

Human leukocyte antigen (HLA) genes provide useful information on the relationship between cancer and the immune system. Despite the ease of obtaining these data through next-generation sequencing methods, interpretation of these relationships remains challenging owing to the complexity of HLA genes. To resolve this issue, we developed a Bayesian method, ALPHLARD-NT, to identify HLA germline and somatic mutations as well as HLA genotypes from whole-exome sequencing (WES) and whole-genome sequencing (WGS) data. ALPHLARD-NT showed 99.2% accuracy for WGS-based HLA genotyping and detected five HLA somatic mutations in 25 colon cancer cases. In addition, ALPHLARD-NT identified 88 HLA somatic mutations, including recurrent mutations and a novel HLA-B type, from WES data of 343 colon adenocarcinoma cases. These results demonstrate the potential of ALPHLARD-NT for conducting an accurate analysis of HLA genes even from low-coverage data sets. This method can become an essential tool for comprehensive analyses of HLA genes from WES and WGS data, helping to advance understanding of immune regulation in cancer as well as providing guidance for novel immunotherapy strategies.

Interval Colorectal Cancer in Inflammatory Bowel Disease: The Role of Guideline Adherence.

Factors associated with interval colorectal cancer (CRC) development in the inflammatory bowel disease (IBD) population remain unclear. Among a cohort of patients with interval CRC, we aimed to evaluate IBD characteristics, colonoscopy quality indicators, and surveillance guideline adherence. We performed a retrospective review of IBD- and non-IBD-associated interval CRCs diagnosed between January 2007 and December 2014 within a large US healthcare system. We evaluated risk factors for CRC among patients with IBD. We assessed adherence to surveillance guidelines according to the American Society for Gastrointestinal Endoscopy (IBD surveillance) and the US Multi-Society Task Force on Colorectal Cancer (polyp surveillance). We compared colonoscopy quality measures between patients with and without IBD. Among 5345 cases of colonic adenocarcinoma, we detected 15 IBD-associated cases of interval CRC and 230 non-IBD-associated cases of interval CRC. Compared to patients without IBD, IBD patients were younger (54.5 vs. 70.4years; p < 0.0001) and experienced a shorter interval between index colonoscopy and CRC diagnosis (20.7 vs. 35.1months; p = 0.0009). Fifty three percent (8/15) of interval CRCs in IBD patients were detected within surveillance guidelines. All IBD patients with interval CRC detected after guideline surveillance interval had high-risk features, including active inflammation, previous low-grade or indefinite dysplasia, multiple pseudopolyps on index colonoscopy, or a first-degree relative with CRC. There were no differences in colonoscopy quality measures between patients with and without IBD. This study stresses the importance of strict short-interval surveillance for IBD patients with high-risk features, including active inflammation on index colonoscopy.

Abstract 2570: Colonic adenocarcinoma-associated DNA-binding site mutations in Rad52 perturbs its ssDNA binding polarity and function in homologous recombination

Abstract Double strand DNA breaks (DSB) are repaired by the Homologous recombination (HR) pathway. Defective HR leads to accumulation of mutations that result in genomic instability and a plethora of associated cancers including, breast, ovarian, and skin cancers. In HR, the DSB is recognized and resected to yield single-stranded DNA (ssDNA) that are immediately coated by replication protein A (RPA). The Rad51 recombinase, which is the central engine for HR, needs to bind to ssDNA buried under RPA and form a nucleoprotein filament. Cells possess mediator proteins that either promote or inhibit the exchange of RPA for Rad51 on ssDNA. RAD52 is one such pro-recombinogenic mediator and functions analogous to the Breast Cancer type 2 Susceptibility Protein (BRCA2) to promote the formation of the Rad51 filament. However, the mechanistic details underlying how Rad52 mediates RPA-Rad51 exchange on ssDNA remains a mystery. Mutations in mediator proteins such as BRCA2 and RAD52 are prevalent in many cancers. For example, the COSMIC database shows there have been 139 sampled cases of mutations of Rad52 in solid tumors. To understand how Rad52 fulfills its mediator functions, it is imperative to first understand how Rad52 associates with DNA. The recent crystal structure of human RAD52 (PDB ID) proposed two ssDNA binding site for RAD52: an outer and an inner binding site, where Arg55 is the interface amino acid between the two sites. To systematically characterize how Rad52 associates with ssDNA, we conducted stopped flow experiments with fluorescently labeled ssDNA. Our results show that Rad52 binds cooperatively to the 5′ end of DNA. Additionally, we created mutations in Rad52-ssDNA binding sites to investigate their role in polarity and cooperativity in depth. These residues and adjacent amino acids are hotspots for mutagenesis in cancers. Abolishing the outer binding site results in a loss of cooperative binding and higher order Rad52-ssDNA complex formation. Mutating the inner binding site abolishes ssDNA binding entirely. Additionally, abolishing the interface arginine residue results in non-selective, rapid binding. This exact amino acid has been mutated in two cases of colonic adenocarcinoma. Our result suggests that both sites are required for maintaining cooperativity and polarity of Rad52 binding to ssDNA. A detailed mechanistic insight into Rad52-ssDNA binding will aid in the development of small-molecule inhibitor drugs that can be used as effective chemotherapeutic agents against cancers in the future. Note: This abstract was not presented at the meeting. Citation Format: Emma A. Tillison, Nilisha Pokhrel, Edwin Antony. Colonic adenocarcinoma-associated DNA-binding site mutations in Rad52 perturbs its ssDNA binding polarity and function in homologous recombination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2570.

Significance of SATB2 expression in colon cancer and its differential diagnosis in digestive tract adenocarcinoma and ovarian primary and metastatic carcinoma

ObjectiveTo study the relationship between the expression of SATB2 in colonic adenocarcinoma and clinicopathological factors, and its role in the identification of ovarian digestive system metastatic adenocarcinoma. MethodsImmunohistochemistry was used to analyze the expression of SATB2 in 130 cases of colon adenocarcinoma, 46 cases of gastric adenocarcinoma, 47 cases of pancreatic ductal adenocarcinoma, 22 cases of cholangiocarcinoma, 53 cases of ovarian mucinous adenocarcinoma, and 38 cases of ovarian metastasis of colorectal carcinoma. Data was analyzed using SPSS 16.0 statistical software. ResultsThe positive expression rate of SATB2 in 130 cases of colon adenocarcinoma was 73.85% (96/130). The expression rate of SATB2 was significantly higher in well -moderately differentiated colon adenocarcinoma than in poorly differentiated adenocarcinoma (p<0.05); the expression rate was significantly higher in cases without tumor deposits than in cases with tumor deposits (p<0.05). The positive expression rate of SATB2 in ovarian metastases of colorectal adenocarcinoma was 81.58% (31/38). No positive expression of SATB2 was observed in other cancers. ConclusionLoss of expression of SATB2 is associated with poor differentiation of primary colon cancer and the formation of tumor deposits. SATB2 provides an ideal diagnostic marker for clinical surgeons to choose the right treatment plan.

Paradoxical effect of regulatory T-cell density in the invasive margin on colon cancer disease-free survival.

47 Background: Increased numbers of FoxP3+ regulatory T cells (Treg), an immunosuppressive T cell subpopulation, in the tumor microenvironment (TME) has been associated with decreased recurrence risk in colon cancer. We assessed the association of Treg levels and disease-free survival (DFS) to confirm correlation with recurrence risk and to evaluate the impact on DFS in patients with colon cancer. Methods: Cases of colonic adenocarcinoma with endoscopic biopsies who had curative resection were collected from 2006-2016. The tissue sections were stained with FoxP3 immunohistochemical stain. FoxP3+ cells (Tregs) were counted with a digital imager software program and reported as cells per square mm. DFS from time of surgery was determined with a retrospective chart review. FoxP3 levels were compared with a Man-Whitney U test and DFS was compared with a Spearman’s test and regression. Results: 109 matched endoscopic biopsy colonic adenocarcinoma specimens were gathered. 94 cases had at least 4 months of follow up and were analyzed. The recurrence rate was 21.3% (20/94) with a median recurrence time of 13.5 months. The median time to follow up in patients with no recurrence was 39 (range 4-116) months. Tregs in those that recurred had a mean density of 152 (range 9-1267) at the invasive margin compared to 251 (range 15-485) in the cases that did not yet recur (p = 0.005). The same association was observed with Treg density in the center of the tumor (p = 0.03). In the cases that had a recurrence, there was a moderate, inverse correlation between Tregs at the invasive margin and time to recurrence (Spearman’s correlation coefficient = -0.46, p = 0.04.) A logarithmic inverse relationship between Tregs at the invasive margin and time to recurrence was observed (Correlation coefficient -0.63, p = 0.003). Conclusions: Treg density in endoscopic biopsies in colon adenocarcinoma is associated with recurrence risk, consistent with previously reported observations. Paradoxically, in patients who did recur, higher Treg density was associated with shorter time to recurrence. This discordant relationship of Treg density and colon cancer prognosis warrants further investigation.

John Cunningham Virus T-Antigen Expression on Mild and Severe Dysplasia Adenomatous Polyp, Low and High Grade Adenocarcinoma of The Colon

Background: John Cunningham Virus (JCV) was involved in pre-malignant lessions and carcinogenesis of the colon. The purpose of this study was to detect and analyze JCV T-Ag expression in mild and severe dysplasia adenomatous polyp as well as low and high grade adenocarcinoma of the colon.Materials and Methods: This study used analytic descriptive, cross sectional approach. The samples’ paraffin blocks were taken from colon adenomatous polyp cases (all grades of dysplasia) and cases of colon adenocarcinoma (all degrees) at Anatomical Pathology Laboratory, School of Medicine, Atma Jaya Catholic University of Indonesia from 2010-2014 (5 years period). Samples were reviewed from HE slides to determine histopathologic diagnosis, grades of dysplasia and grading. We performed immunohistochemistry staining with monoclonal antibody anti–SV 40-T-Ag to detect JCV T-Ag expression.Results: We found 7 cases of colon adenomatous polyp, of which 4 (57%) were mild dysplasia and 3 (43%) were severe dysplasia. Positive expression of JCV T-Ag was detected in 1 (14%) mild dysplasia case. Data analysis using Fischer’s Exact Test was p&gt;0.05. We also found 16 cases of colon adenocarcinoma. 14 cases (87.5%) of low grade variant and 2 cases (12.5%) of high grade variant. Positive expression of JCV T-Ag was detected in 2 (12.5%) low grade cases. Data analysis using Fischer’s Exact Test was p&gt;0.05.Conclusion: There was no difference of JCV T-Ag expression in colon adenomatous polyp (mild-severe dysplasia) and colon adenocarcinoma (low-high grade) cases at Anatomical Pathology Laboratory School of Medicine, Atma Jaya Catholic University of Indonesia 2010-2014.Keywords: colon adenomatous polyp, adenocarcinoma, JCV T-Ag

Competitive Endogenous RNA (ceRNA) Regulation Network of lncRNA-miRNA-mRNA in Colorectal Carcinogenesis.

Competitive endogenous RNA (ceRNA) regulation suggested complex network of all transcript RNAs including long noncoding RNAs (lncRNAs), which can act as natural miRNA sponges to inhibit miRNA functions and modulate mRNA expression. Until now, the specific ceRNA regulatory mechanism of lncRNA-miRNA-mRNA in colorectal cancer (CRC) still remains unclear. RNA sequencing data of 478 colon adenocarcinoma cases and 41 controls as well as 166 rectum adenocarcinoma cases and 10 controls were obtained from The Cancer Genome Atlas (TCGA) to investigate the significant changes of lncRNAs, miRNAs and mRNAs in colorectal carcinogenesis. The target lncRNAs and mRNAs of miRNAs were predicted by miRWalk. Functional and enrichment analyses were conducted by DAVID database. The lncRNA-miRNA-mRNA interaction network was constructed using Cytoscape. We constructed ceRNA regulatory networks including 22 up-regulated lncRNAs, 12 down-regulated miRNAs and 122 up-regulated mRNAs, as well as 8 down-regulated lncRNAs, 43 up-regulated miRNAs and 139 down-regulated mRNAs. The GO enrichment showed that up-regulated genes mainly enriched in biological process including organic anion transport, collagen catabolic process, wound healing, Wnt receptor signalling and in pathways of tyrosine metabolism, taurine and hypotaurine metabolism, melanogenesis and phenylalanine metabolism. For down-regulated genes, significant enrichment was found in biological process of metal ion homeostasis, transmission of nerve impulse, cell-cell signalling, transmembrane transport and in pathways of ABC transporters, neuroactive ligand-receptor interaction, retinol metabolism, nitrogen metabolism and steroid hormone biosynthesis. We identified significantly altered lncRNAs, miRNAs and mRNAs in colorectal carcinogenesis, which might serve as potential biomarkers for tumorigenesis of CRC. In addition, the ceRNA regulatory network of lncRNA-miRNA-mRNA was constructed, which would elucidate novel molecular mechanisms involved in initiation and progression of CRC, thus providing promising clues for clinical diagnosis and therapy.

Assessment of Relationship Between Expression of Survivin Protein and Histopathology Diagnosis and Malignancy Severity in Colon Specimen.

Survivin is a member of the inhibitor of an apoptosis protein family that has been shown to inhibit apoptosis, promote cell proliferation and enhance angiogenesis. In this study, the survivin protein expression in normal, colon polyp, and adenocarcinoma tissues was investigated. Immunohistochemical staining for nuclear survivin was carried out on 45 normal colon tissue samples, 38 samples of a colonic polyp, and 37 cases of colon adenocarcinoma operated by colonoscopy or colectomy. The percentages of cells that expressed survivin were classified qualitatively into four categories (0, 1+, 2+, and 3+) based on the intensity of staining and the percentage of cells. An area of samples with colon polyp diagnosis or colon adenocarcinoma that had no microscopic pathology was considered as normal tissues. Survivin protein expression was negative in all cases of normal colon tissue samples while it was expressed in 31 out of 38 colon polyp specimens (81.5%) and in 35 out of 37 (94.5%) colon adenocarcinoma samples. Amount of expression in the colon adenocarcinoma (p < 0.001) was significantly higher than the amount of expression in the colon polyp. There was not a significant correlation between the survivin protein expression and the low and high grade adenocarcinoma (p = 0.874). Survivin protein was not expressed in normal colon tissues and its amount was higher in the colonic adenocarcinoma compared to the colon polyp. Due to the variations in the intensity of expression in colon polyp (changing from negative to + 3), this marker cannot be used for differentiating the polyp from the adenocarcinoma.

A Rare Case of Metastatic Colon Cancer Involving the Right Atrium Presenting as Superior Vena Cava Syndrome

Colorectal cancer is one of the most common forms of malignancies and is well documented to metastasize to liver, lungs and peritoneum. Only a handful of well-documented cases of colorectal cancer metastasis to the right atrium have been documented. As such, we report a rare case of colon adenocarcinoma that metastasized to the right atrium and resulted in superior vena cava (SVC) syndrome. The patient is a 50-year-old male with history of hyperlipidemia, anxiety and metastatic colon cancer involving lungs and liver previously treated with chemotherapy and recently with immunotherapy who was transferred to our hospital for management of occluded SVC with the mass extending into the left atrium. The patient initially presented at another hospital for facial and upper extremity swelling and shortness of breath. A CT of the chest noted occluded SVC with a right atrial mass. This was further confirmed on transesophageal echocardiography which noted a large mass occupying most of the right atrial cavity measuring 6.8cm x 4.6cm and was noted to prolapse across the tricuspid valve in to the right ventricle. He was started on Eliquis and was transferred to our hospital for further management. He underwent an angiovac thrombectomy with Interventional Radiology (IR). During the procedure, findings of right atrial mass extending into the SVC were noted with subsequent partial debulking of the mass. Post procedure, the patient reported moderated improvement of his symptoms of dyspnea and swelling. Microscopic pathologic examination of the excised mass exhibited histomorphological features of metastatic colorectal cancer as cells demonstrated strong and diffuse nuclear reactivity for CDX2 stain. CT surgery evaluated patient for further surgical evaluation of right atrial mass, however he was deemed a poor surgical candidate. As such, IR placed a stent in the SVC to further alleviate SVC occlusion. Prior to discharge, Oncology also evaluated patient and recommend continued palliative radiation as outpatient. Conclusion: Although rare, any patient with a history of colorectal adenocarcinoma who presents with cardiopulmonary symptoms and suspicious cardiac mass should undergo evaluation for possible cardiac metastasis. It is recommended that a multidisciplinary approach involving cardiologists, oncologists, IR, and cardiothoracic surgeons is essential for determining the optimal therapeutic regimen for such patients.1483_A Figure 1. CT of the chest exhibiting the Right atrial mass1483_B Figure 2. Right atrial mass and thrombus extending into superior vena cava1483_C Figure 3. Microscopic pathologic examination of the excised mass exhibited histomorphological features of metastatic colorectal cancer as cells demonstrated strong and diffuse nuclear reactivity for CDX2 stain

A Rare Case of Colonic Adenocarcinoma with Mucinous Features

A Rare Case of Colonic Adenocarcinoma with Mucinous Features

Diagnostic Utility of PD-L1 Expression in Lung Adenocarcinoma: Immunohistochemistry and RNA In Situ Hybridization

Programmed death receptor and programmed death ligand (PD-L1) are immunoregulatory proteins. Nonsmall cell lung cancer bypasses the immune system through the induction of protumorigenic immunosuppressive changes. The better understanding of immunology and antitumor immune responses has brought the promising development of novel immunotherapy agents like programmed death receptor checkpoint inhibitors. The aim of this study was to investigate the expression of PD-L1 in lung adenocarcinoma (ADC), comparing 2 different technologies: immunohistochemistry (IHC) by 2 methods versus RNA in situ hybridization (RISH). In total, 20 cases of ADC of the lung and 4 samples of metastatic colon ADC were selected. Evaluation of PD-L1 expression was performed by IHC and RISH. RISH was performed using RNAscope. Both methods were scored in tumor cells and quantified using combined intensity and proportion scores. Eight of 20 (40%) lung ADC and 2 of 4 (50%) colon ADC were positive for PD-L1 with Cell Signaling IHC, and 65% lung ADC were positive by Dako IHC (13/20). All 4 cases of colon ADC were negative. When evaluated by RISH, 12 lung ADC (60%) and 1 colon ADC (25%) were PD-L1 positive. RNAscope probes provide sensitive and specific detection of PD-L1 in lung ADC. Both IHC methods (Cell Signaling and Dako) show PD-L1 expression, with the Dako method more sensitive (40% vs. 65%). This study illustrates the utility of RISH and Cell Signaling IHC as complementary diagnostic tests, and Food and Drug Administration approved Dako IHC as a companion diagnostic test.

MLH1-rheMac hereditary nonpolyposis colorectal cancer syndrome in rhesus macaques

Over the past two decades, 33 cases of colonic adenocarcinomas have been diagnosed in rhesus macaques (Macaca mulatta) at the nonhuman primate colony of the Keeling Center for Comparative Medicine and Research at The University of Texas MD Anderson Cancer Center. The distinctive feature in these cases, based on PET/computed tomography (CT) imaging, was the presence of two or three tumor lesions in different locations, including proximal to the ileocecal juncture, proximal to the hepatic flexure, and/or in the sigmoid colon. These colon carcinoma lesions selectively accumulated [18F]fluorodeoxyglucose ([18F]FDG) and [18F]fluoroacetate ([18F]FACE) at high levels, reflecting elevated carbohydrate and fatty acid metabolism in these tumors. In contrast, the accumulation of [18F]fluorothymidine ([18F]FLT) was less significant, reflecting slow proliferative activity in these tumors. The diagnoses of colon carcinomas were confirmed by endoscopy. The expression of MLH1, MSH2, and MSH6 proteins and the degree of microsatellite instability (MSI) was assessed in colon carcinomas. The loss of MLH1 protein expression was observed in all tumors and was associated with a deletion mutation in the MLH1 promoter region and/or multiple single-nucleotide polymorphism (SNP) mutations in the MLH1 gene. All tumors exhibited various degrees of MSI. The pedigree analysis of this rhesus macaque population revealed several clusters of affected animals related to each other over several generations, suggesting an autosomal dominant transmission of susceptibility for colon cancer. The newly discovered hereditary nonpolyposis colorectal cancer syndrome in rhesus macaques, termed MLH1-rheMac, may serve as a model for development of novel approaches to diagnosis and therapy of Lynch syndrome in humans.

Un cas d’adénocarcinome colique « trèfle à quatre feuilles »

Un cas d’adénocarcinome colique « trèfle à quatre feuilles »

Morphometric analysis and immunohistochemical expression of cytochrome C oxidase in colonic adenomas and adenocarcinomas

Background/aim Colorectal cancer is the seventh most common cancer in Egypt, constituting 3.47% of male cancers and 3% of female cancers. The aim of the current study is to evaluate the immunohistochemical expression of cytochrome C oxidase (CcO) as well as the nuclear morphometric measurements in colonic adenomas and adenocarcinomas. Materials and methods Sixty cases of colonic biopsies/specimens were involved in this study. The samples were received in the Department of Pathology of Kasr El-Aini Hospital, Cairo University, Egypt, and they were divided into three groups: 20 cases of normal control, 20 cases of colonic adenoma, and 20 cases of colonic adenocarcinoma. The expression of CcO was studied by immunohistochemistry in 60 formalin-fixed paraffin blocks. Nuclear morphometric parameters were evaluated using an image analysis system. The area percentage of the marker expression was correlated with nuclear morphometric parameters. Results Immunohistochemical study showed that 65% of control biopsies showed a significant difference with colonic adenocarcinoma cases in contrast and adenoma cases showed a nonsignificant difference with normal control biopsies. Mean areas percentages of CcO expression were significantly different between all three studied groups. The means of nuclear area, length, and perimeter showed a significant difference while means nuclear width and roundness showed a nonsignificant difference between all three studied groups. There was significant correlation between mean area percentage of CcO expression with mean nuclear area and perimeter in control, adenoma, and adenocarcinoma cases. Conclusion CcO protein is significantly expressed in the cytoplasm of normal colonic mucosa. It was significantly deficient in colonic adenocarcinoma. Nuclear morphometry can be introduced as a useful morphologic feature of patients with variable colonic lesions specially in combination with CcO.

Alternative splicing events implicated in carcinogenesis and prognosis of colorectal cancer.

Background: Emerging evidence suggested that aberrant alternative splicing (AS) is pervasive event in development and progression of cancer. However, the information of aberrant splicing events involved in colorectal carcinogenesis and progression is still elusive.Materials and Methods: In this study, splicing data of 499 colon adenocarcinoma cases (COAD) and 176 rectum adenocarcinoma (READ) with clinicopathological information were obtained from The Cancer Genome Atlas (TCGA) to explore the changes of alternative splicing events in relation to the carcinogenesis and prognosis of colorectal cancer (CRC). Gene interaction network construction, functional and pathway enrichment analysis were performed by multiple bioinformatics tools.Results: Overall, most AS patterns were more active in CRC tissues than adjacent normal ones. We detected altogether 35391 AS events of 9084 genes in COAD and 34900 AS events of 9032 genes in READ, some of which were differentially spliced between cancer tissues and normal tissues including genes of SULT1A2, CALD1, DTNA, COL12A1 and TTLL12. Differentially spliced genes were enriched in biological process including muscle organ development, cytoskeleton organization, actin cytoskeleton organization, biological adhesion, and cell adhesion. The integrated predictor model of COAD showed an AUC of 0.805 (sensitivity: 0.734; specificity: 0.756) while READ predictor had an AUC of 0.738 (sensitivity: 0.614; specificity: 0.900). In addition, a number of prognosis-associated AS events were discovered, including genes of PSMD2, NOL8, ALDH4A1, SLC10A7 and PPAT.Conclusion: We draw comprehensive profiles of alternative splicing events in the carcinogenesis and prognosis of CRC. The interaction network and functional connections were constructed to elucidate the underlying mechanisms of alternative splicing in CRC.

Lenalidomide in combination with R‐ESHAP in patients with relapsed or refractory diffuse large B‐cell lymphoma: A phase 2 study from the Spanish group GELTAMO

Introduction: Diffuse large B-cell lymphoma (DLBCL) patients with relapsed or refractory disease after rituximab-containing therapy have poor outcomes with current salvage regimens. We conducted a phase 1b/2 trial to analyze the safety and efficacy of adding lenalidomide to R-ESHAP regimen (LR-ESHAP). The phase 1b part of the trial has been completed, and lenalidomide 10 mg/day was identified as the maximum tolerated dose (Br J Haematol 2016; 173: 245-52). Here we present the preliminary results of the phase 2 part (ClinicalTrials.gov Identifier: NCT02340936). Methods: Eligible patients must have relapsed or refractory DLBCL after first-line treatment with rituximab combined with an anthracycline-containing regimen. Subjects received 3 cycles of lenalidomide 10 mg given on days 1 to 14 of every 21-day cycle, in combination with R-ESHAP at standard doses. Responding patients received BEAM conditioning followed by autologous stem-cell transplantation (ASCT). The primary end point was overall response rate (ORR) after 3 cycles of therapy. Secondary end points were complete remission (CR) rate, stem-cell mobilization activity, progression-free survival (PFS), overall survival (OS), and toxicity. Results: A total of 46 patients (median age 58 [23-69] years, 56.5% male) were included (January 2012-November 2015). Evaluable population per-protocol consisted of 44 patients. Forty-two serious adverse events were reported during 128 cycles of LR-ESHAP, including 14 episodes of febrile neutropenia (11%), 12 infections (9%), 2 renal disorders, 3 cardiac disorders, 3 thrombosis, and 8 other toxicities, all of them recovered except 1 case of colon adenocarcinoma. There were no treatment-related deaths. Forty out of 44 patients (91%) received the planned 3 cycles of treatment (2 without lenalidomide in the third cycle). ORR to LR-ESHAP was 68% (41% CR). Patients with relapsed disease had significantly better ORR and CR rates (87% and 67%, respectively) than patients in PR after first-line (77% and 31%) or with primary refractory disease (<PR) (37% and 25%) (p = 0.004 and 0.02, respectively). Evaluation of response according to cell of origin (COO) is ongoing. So far, 30 and 21 patients, respectively, have been analyzed according to Hans' algorithm (ORR of 62% and 44% in germinal-center B-cell-like [GCB] [n = 21] and non-GCB [n = 9] groups, respectively, p > 0.1) and gene expression profiling (ORR of 71%, and 75% in GCB [n = 14], and activated B-cell-like [ABC] [n = 4] groups, respectively, p > 0.1). Thirty-nine out of 42 patients (93%) were successfully mobilized after one (n = 32) or 2 (n = 7) mobilization procedures, and 28 (64% of evaluable patients) underwent ASCT according to protocol, one of them after bone-marrow harvest. With a median follow-up of 15 months (11-53), the estimated 2-year PFS and OS were 54% and 52%, respectively. Conclusions: LR-ESHAP is safe, feasible, and associated with high-response rates in rituximab-pretreated relapsed or refractory DLBCL patients. Efficacy analysis according to COO and other biological factors is ongoing. Keywords: diffuse large B-cell lymphoma (DLBCL); lenalidomide; salvage treatment

Differential expression of nucleolin in colon adenoma and adenocarcinoma

Abstract Objective The aim of the study was to investigate the discrepancy in nucleolin expression between colon adenoma and colon adenocarcinoma, explore the role of nucleolin expression in the carcinogenesis of colon adenocarcinoma, and determine the correlation of the nucleolin expression level with histological grade in colon adenocarcinoma. Methods In total, 80 cases of colon adenocarcinoma with cancer-adjacent colon mucosa and 60 cases of colon adenomas were examined by immunohistochemistry using an antibody against nucleolin. Nucleolin expression levels in these groups were compared. The correlation between the nucleolin expression level and grade of colon adenocarcinoma was analyzed. Results Nucleolin expression is located in the nuclei of colon adenocarcinoma, colon adenoma, and cancer-adjacent colon mucosa tissues with different intensities. A semiquantitative evaluation using the Allred scoring system showed that the nucleolin immunostaining score in colon adenocarcinoma (7.8 ± 0.1) was significantly higher than those in colon adenoma (6.3 ± 0.2) and cancer-adjacent colon mucosa (5.4 ± 0.1; P &lt; 0.01). The nucleolin immunostaining score in colon adenoma was significantly higher than that in cancer-adjacent colon mucosa (P &lt; 0.01). Nucleolin expression levels in well-differentiated and moderately differentiated adenocarcinoma (6.8 ± 0.2) were significantly lower than those in poorly differentiated adenocarcinoma (8.0 ± 0.1; P &lt; 0.01). Conclusion Increased nucleolin expression may play an important role in the process of malignant transformation of colon adenocarcinoma and predicts a poor prognosis.

Lenalidomide in Combination with R-ESHAP (LR-ESHAP) in Patients with Relapsed or Refractory Diffuse Large B-Cell Lymphoma Who Are Candidates for Autologous Stem-Cell Transplantation: A Phase 2 Study from the Spanish Group Geltamo

Background: Diffuse large B-cell lymphoma (DLBCL) patients with relapsed or refractory disease after rituximab-containing first-line therapy have poor outcomes with current salvage regimens. Accordingly, prospective studies incorporating new agents are needed for these patients. We conducted a phase 1b/2 trial to analyze the safety and efficacy of lenalidomide in combination with R-ESHAP (LR-ESHAP) in patients with relapsed or refractory DLBCL. The phase 1b part of the trial has been completed and lenalidomide 10 mg/day was identified as the maximum tolerated dose (MTD) (Martín et al, Br J Haematol 2016; 173: 245-52). Here we present the preliminary results of the phase 2 (ClinicalTrials.gov Identifier: NCT02340936).Patients and methods: Eligible patients must be refractory to, or have relapsed following first-line treatment with rituximab in combination with an anthracycline-containing regimen and be eligible for autologous stem-cell transplantation (ASCT). Subjects received 3 cycles of lenalidomide 10 mg given on days 1 to 14 of every 21-day cycle, in combination with R-ESHAP salvage chemotherapy at standard doses (rituximab 375 mg/m2 day 1, etoposide 40 mg/m2 days 1-4, cisplatin 25 mg/m2 days 1-4, cytarabine 2000 mg/m2 day 5, and methylprednisolone 500 mg days 1-5). Responding patients received BEAM followed by ASCT. The primary endpoint was overall response rate (ORR) after 3 cycles of therapy. Secondary endpoints were complete remission (CR) rate, stem-cell mobilization activity, progression-free and overall survival, and toxicity.Results:Patient characteristics: 46 patients were included in the phase 2 analysis, of whom 12 were enrolled between January 2012 and March 2013, and had been given the MTD during phase 1; 34 were enrolled between January 2015 and November 2015. Median age was 58 (23-69) years, and 56.5% were male. Evaluable population per-protocol consisted of 44 patients because 2 patients were excluded during the first cycle due to withdrawal of consent and centralized diagnosis at relapse of lymphoblastic lymphoma, respectively. First-line treatment consisted of R-CHOP or similar in 38 patients, R-EPOCH in 3, VR-CAP in 1, and Burkitt lymphoma protocols in 2. Disease status at LR-ESHAP was: primary refractory disease in 67% of patients (partial response [PR] after first-line, n=13; and stable or progressive disease [<PR] after first-line, n=16), and relapsed disease in 33% (5 relapses occurred <1 year from diagnosis, and 10 late relapses). IPI was 0-1 in 38%, 2-3 in 48%, and 4-5 in 14%.Feasibility and efficacy: 40 out of 44 patients (91%) received the planned 3 cycles of treatment (2 without lenalidomide in the third cycle due to toxicity) and 2 patients two cycles (due to persistent hematological and renal toxicities, respectively). Two patients discontinued treatment during the first cycle due to grade 3 infections (fungal pulmonary infection and mastoiditis, respectively). ORR to LR-ESHAP in the per-protocol population (n=44) was 68% (41% CR). Patients with relapsed disease had significantly better ORR and CR rates (87% and 67%, respectively) than patients in PR (77% and 31%) or <PR (37% and 25%) (p=0.004 and 0.02, respectively). Evaluation of response according to cell of origin (COO) is ongoing. Thirty-nine out of 42 patients (93%) were successfully mobilized after one (n=32) or two (n=7) mobilization procedures, and 28 (64%) underwent ASCT according to protocol, 1 of them after bone-marrow harvest. Reasons for not performing the transplant were: early progression (n=13), trial discontinuation due to toxicity (n=2), mobilization failure (n=1), and diagnosis of colon adenocarcinoma (n=1).Toxicity: 42 serious adverse events (SAEs) have been reported during 128 cycles of treatment, including 14 episodes of febrile neutropenia (11%), 12 infections (9%), 2 renal disorders, 3 cardiac disorders, 3 thrombosis, and 8 other toxicities. All of them recovered except 1 case of colon adenocarcinoma in a patient with previous adenoma in the same location. There were no treatment-related deaths during LR-ESHAP cycles.Conclusions: LR-ESHAP is safe, feasible and associated with high response rates in rituximab-pretreated relapsed or refractory DLBCL patients. Longer follow-up is needed to perform survival analysis. Efficacy analysis according to COO is ongoing. DisclosuresMartín:Janssen: Honoraria; Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Sevier: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Gilead: Membership on an entity’s Board of Directors or advisory committees. González-Barca:Sanofi: Membership on an entity’s Board of Directors or advisory committees; Servier: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Gilead: Honoraria; Janssen: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Roche: Honoraria. Lopez-Guillermo:Roche: Consultancy, Research Funding; Celgene: Consultancy; Gilead: Consultancy; Janssen: Consultancy; Mundipharma: Consultancy.

E. coli Sepsis: Red Flag for Colon Carcinoma

Introduction: While the relationship between Streptococcus bovis bacteremia and colon cancer has been well established, rare cases of colon adenocarcinoma have been reported after exposure to other microorganisms. We present an atypical case of newly diagnosed colon cancer and provide insight into the infectious predispositions of E. coli bacteremia to the development of colon adenocarcinoma. Case Report: A 66-year-old female with past medical history of hypertension, type 2 diabetes mellitus, and obesity was evaluated for one week symptoms of subjective fevers, chills and lower back pain. Upon initial evaluation, her temperature was 101 degrees Fahrenheit with a white count of 12,000 K/mm3, procalcitonin level of 29.02 K/mm3, and lactic acid of 4.8 MMOL/L. Initial septic workup was positive for E. coli bacteremia. The patient was started on Aztreonam (due to her Penicillin allergy). Repeat blood culture 48 hours later on antibiotics was negative for any growth. However, later during hospital stay blood culture was repeated due to SIRS, which was positive again for E. coli. The patient reported no abdominal complaints other than chronic constipation. The patient had cholecystectomy done many years ago. No family history of colorectal cancer and never had a colonoscopy. CT scan of the chest/abdomen/pelvis with contrast revealed no signs of colitis however; it was remarkable for spondylitis/diskitis with adjacent fluid collection at L4-L5. The patient underwent a CT guided aspiration biopsy of the L4-L5 discitis and adjacent fluid collection, showed no growth on culture. Without clear etiology for recurrent E. coli bacteremia ultimately colonoscopy was performed which showed an ulcerated mass in the cecum. Biopsy showed moderately differentiated adenocarcinoma. Discussion: CRC presenting as a persistent E. coli bacteremia is rare. Molecular research has supported the relationship between the virulent E. coli strains and the development of CRC. E. coli strains B2 and D produce cyclomodulin toxins as part of their virulence, which interfere with the cell cycle regulation, promoting chromosomal instability, and increasing susceptibility to the cancer. Enteropathogenic E. coli was reported to down regulate expression of key DNA mismatch repair proteins MSH2 and MLH1. hly+ type 1- E. coli is also associated with adenoma and CRC development. The patients with recurrent E. coli bacteremia with an unknown source, colonoscopy should be done to look for colon tumor.