Villous Growth Research Articles

Percutaneous transhepatic cholangioscopy-assisted biliary polypectomy for local palliative treatment of intraductal papillary neoplasm of the bile duct.

Intraductal papillary neoplasm of the bile duct (IPNB) is a premalignant biliary-type epithelial neoplasm with intraductal papillary or villous growth. Currently reported local palliative therapeutic modalities, including endoscopic nasobiliary drainage, stenting and biliary curettage, endoscopic biliary polypectomy, percutaneous biliary drainage, laser ablation, argon plasma coagulation, photodynamic therapy, and radiofrequency ablation to relieve mechanical obstruction are limited with weaknesses and disadvantages. We have applied percutaneous transhepatic cholangioscopy (PTCS)-assisted biliary polypectomy (PTCS-BP) technique for the management of IPNB including mucin-hypersecreting cast-like and polypoid type tumors since 2010. To assess the technical feasibility, efficacy, and safety of PTCS-BP for local palliative treatment of IPNB. Patients with mucin-hypersecreting cast-like or polypoid type IPNB and receiving PTCS-BP between September 2010 and December 2019 were included. PTCS-BP was performed by using a half-moon type snare with a soft stainless-steel wire, and the tumor was snared and resected with electrocautery. The primary outcome was its feasibility, indicated by technical success. The secondary outcomes were efficacy, including therapeutic success, curative resection, and clinical success, and safety. Five patients (four with mucin-hypersecreting cast-like type and one with polypoid type IPNB) were included. Low- and high-grade intraepithelial neoplasia (HGIN) and recurrent IPNB with invasive carcinoma were observed in one, two, and two patients, respectively. Repeated cholangitis and/or obstructive jaundice were presented in all four patients with mucin-hypersecreting cast-like type IPNB. All five patients achieved technical success of PTCS-BP. Four patients (three with mucin-hypersecreting cast-like type and one with polypoid type IPNB) obtained therapeutic success; one with mucin-hypersecreting cast-like type tumors in the intrahepatic small bile duct and HGIN had residual tumors. All four patients with mucin-hypersecreting IPNB achieved clinical success. The patient with polypoid type IPNB achieved curative resection. There were no PTCS-BP-related serious adverse events. PTCS-BP appears to be feasible, efficacious, and safe for local palliative treatment of both mucin-hypersecreting cast-like and polypoid type IPNB.

Abstract 4769: Phospho-YAP (Ser127) overexpression is associated with sporadic advanced adenomas: The Tennessee Colorectal Polyp Study

Abstract About 70-80% of sporadic colorectal cancers (CRCs) harbor APC mutations. Whether specific characteristics of APC mutation or APC-related signaling pathways contribute to tumor initiation, advanced progression, and recurrence in the precancerous lesions remain to be elucidated. This study included145 sporadic precancerous adenomas from the Tennessee Colorectal Polyp Study. Expression of six APC-related proteins (β-catenin, p-YAP, CtBP, EB1, Asef, and N-terminal APC) in adenoma and non-tumor tissues (n=22) were quantitatively detected by immunohistochemistry. Targeted sequencing of APC was done for 108 adenomas. Metachronous adenoma status was obtained from chart review. YAP-correlated genes and their functional enrichment analysis were conducted using gene expression data of 326 adenomas from the Gene Expression Omnibus database. We found that nuclear β-catenin and p-YAP overexpression only existed in adenoma tissue, correlated with each other (r=0.26, P=0.0018), and associated with APC mutations (P=0.05). P-YAP was strongly associated with odds of large adenomas (OR=14.25, 95%CI=3.32-61.18, P trend<0.0001), villous growth pattern (OR=58.71, 95%CI=10.49-328.61, P trend<0.0001) and advanced adenomas (OR=12.62, 95%CI 4.57-34.86, P trend<0.001), and showed a significant interaction with nuclear β-catenin on advanced tumor (OR=16.82, 95%CI 4.41-64.08, P<0.0001). After adjusting for selected covariates and mutually adjusting for biomarkers with P<0.20 in crude models, p-YAP overexpression remained significantly associated with advanced adenomas (OR=12.31, 95% CI=3.78-40.10, P trend<0.0001). YAP-correlated genes were significantly enriched in autophagy, unfolded protein response, and sirtuin pathways with predominantly pro-tumorigenic alterations. APC mutation and selected biomarkers were not significantly associated with metachronous adenoma. In conclusion, p-YAP overexpression might be a promising biomarker for high-risk sporadic adenoma. In addition to its synergistic effect with nuclear β-catenin, p-YAP may play tumorigenic roles through interaction with other cancer pathways in human sporadic adenoma. This study provided new evidence for oncogenic effects of p-YAP in human sporadic adenomas and shed light on the complex network of Hippo, Wnt, and other oncogenic signaling pathways. Citation Format: Lei Fan, Xingyi Guo, Mary Kay Washington, Jiajun Shi, Reid M. Ness, Qi Liu, Wanqing Wen, Xiao Liu, Qiuyin Cai, Wei Zheng, Robert J. Coffey, Martha J. Shrubsole, Timothy Su. Phospho-YAP (Ser127) overexpression is associated with sporadic advanced adenomas: The Tennessee Colorectal Polyp Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4769.

A Comparison of Risk Classification Systems of Colorectal Adenomas: A Case-Cohort Study

Because post-polypectomy surveillance uses a growing proportion of colonoscopy capacity, more targeted surveillance is warranted. We therefore compared surveillance burden and cancer detection using 3 different adenoma classification systems. In a case-cohort study among individuals who had adenomas removed between 1993 and 2007, we included 675 individuals with colorectal cancer (cases) diagnosed a median of 5.6 years after adenoma removal and 906 randomly selected individuals (subcohort). We compared colorectal cancer incidence among high- and low-risk individuals defined according to the traditional (high-risk: diameter ≥10 mm, high-grade dysplasia, villous growth pattern, or 3 or more adenomas), European Society of Gastrointestinal Endoscopy (ESGE) 2020 (high-risk: diameter ≥10 mm, high-grade dysplasia, or 5 or more adenomas), and novel (high-risk: diameter ≥20 mm or high-grade dysplasia) classification systems. For the different classification systems, we calculated the number of individuals recommended frequent surveillance colonoscopy and estimated number of delayed cancer diagnoses. Four hundred and thirty individuals with adenomas (52.7%) were high risk based on the traditional classification, 369 (45.2%) were high risk based on the ESGE 2020 classification, and 220 (27.0%) were high risk based on the novel classification. Using the traditional, ESGE 2020, and novel classifications, the colorectal cancer incidences per 100,000 person-years were 479, 552, and 690 among high-risk individuals, and 123, 124, and 179 among low-risk individuals, respectively. Compared with the traditional classification, the number of individuals who needed frequent surveillance was reduced by 13.9% and 44.2%, respectively, and 1 (3.4%) and 7 (24.1%) cancer diagnoses were delayed using the ESGE 2020 and novel classifications. Using the ESGE 2020 and novel risk classifications will substantially reduce resources needed for colonoscopy surveillance after adenoma removal.

Intraductal oncocytic papillary neoplasm arising in Peutz-Jeghers Syndrome bile duct: a unique case report

BackgroundPeutz-Jeghers syndrome (PJS) is a rare, autosomal dominant disorder caused by germline mutations of STK11/LKB1, with an increased risk of tumors at multiple sites. Intraductal oncocytic papillary neoplasm (IOPN) is a unique subtype of intraductal papillary neoplasm of the bile duct (IPNB) defined by a premalignant neoplasm with intraductal papillary or villous growth of biliary-type epithelium. IOPN has a distinct mutation profile compared with both IPNB and intraductal papillary mucinous neoplasm (IPMN).Case presentationWe herein describe the case of a 44-year-old woman who presented as polyps in the intestinal lumen of sigmoid colon and a 3.1 × 2.1 cm mass in the left lobe of liver. Gross feature revealed a cystic papillary mass and the neoplasm had a clear boundary with the surrounding liver tissue. Histology revealed complex papillary structures, a small amount of fine fibrovascular cores and immunohistochemistry showed extensive positive for MUC5AC, MUC6, CD117. Therefore, histological and immunohistochemical examination of the liver tumor suggested the diagnosis of IOPN. Next-generation sequencing (NGS) revealed other than STK11 germline mutation, the tumor also harbors GNAS somatic mutation at codon 478 and EGFR amplification.ConclusionTo our knowledge, this is the first report of IOPN arising in PJS. This case enlarges the spectrum of PJS related tumors and genetic rearrangements in IOPN.

High-Grade Appendiceal Mucinous Neoplasm: Clinicopathologic Findings in 35 Cases.

High-grade appendiceal mucinous neoplasm (HAMN) is a relatively recently introduced term describing a rare epithelial neoplasm of the appendix that demonstrates pushing-type invasion but high-grade cytologic atypia. It remains understudied. To describe clinicopathologic features of HAMNs. We identified 35 HAMNs in a multi-institutional retrospective study. Clinical and histologic features were reviewed in all cases, as well as molecular features in 8 cases. Patients were 57 years of age on average and most commonly presented with abdominal/pelvic pain. Histologically, 57% of the tumors showed widespread high-grade features. Architectural patterns in high-grade areas included flat, undulating, or villous growth, and occasionally micropapillary, cribriform, or multilayered growth. Thirteen cases had intact serosa, and the remaining 22 perforated the serosa, including 7 with peritoneal acellular mucin beyond appendiceal serosa and 10 with grade 2 pseudomyxoma peritonei. Molecular abnormalities included KRAS mutations in 7 cases and TP53 mutations in 4. No tumor confined to the appendix recurred. Two patients without pseudomyxoma peritonei at initial presentation developed pseudomyxoma on follow-up. Among 11 patients who presented with pseudomyxoma peritonei, 5 died of disease and 3 were alive with disease at last follow-up. HAMNs have a similar presentation to low-grade appendiceal mucinous neoplasm, and similar stage-based prognosis. When they spread to the peritoneum, they typically produce grade 2 pseudomyxoma peritonei, which may be associated with a worse prognosis than classical grade 1 pseudomyxoma peritonei.

CircHIPK3 contributes to human villous trophoblast growth, migration and invasion via modulating the pathway of miR-346/KCMF1.

Preeclampsia (PE) is one of the leading causes of maternal and perinatal morbidity and mortality worldwide. The regular growth, migration and invasion of villous trophoblast cells contribute to placental development. The objective of this study was to investigate the role and mechanism of circular RNA homeodomain interacting protein kinase 3 (circHIPK3) in the biological functions of trophoblast cells. The expression of circHIPK3, microRNA-346 (miR-346) and potassium channel modulatory factor 1 (KCMF1) mRNA was measured by quantitative real-time PCR (qPCR). Trophoblast cell proliferation, migration/invasion and cell cycle progression/apoptosis were determined by CCK-8 assay, transwell assay and flow cytometry assay, respectively. The predicted relationship between miR-346 and circHIPK3 or KCMF1 by bioinformatics was confirmed dual-luciferase reporter assay and RIP assay. CircHIPK3 and KCMF1 were downregulated, while miR-346 was upregulated in placenta tissues from PE patients. The forced expression of circHIPK3 promoted trophoblast cell proliferation and migration/invasion but alleviated cell cycle arrest and cell apoptosis. MiR-346 was a target of circHIPK3, and miR-346 restoration reversed the effects of circHIPK3 upregulation. In addition, circHIPK3 acted as miR-346 sponge to modulate KCMF1 expression. KCMF1 downregulation partially repressed trophoblast cell proliferation, migration and invasion that were facilitated by miR-346 inhibition or circHIPK3 upregulation. CircHIPK3 contributes to trophoblast cell proliferation, migration and invasion by upregulating KCMF1 via acting as miR-346 sponge.

Monocyte Chemotactic Proteins (MCP) in Colorectal Adenomas Are Differently Expressed at the Transcriptional and Protein Levels: Implications for Colorectal Cancer Prevention.

The expression of monocyte chemotactic proteins (MCPs) in colorectal polyps and their suitability as targets for chemoprevention is unknown, although MCP expression and secretion can be modulated by non-steroidal inflammatory drugs. This study was designed to determine the expression patterns of MCP-1/CCL2, MCP-2/CCL8, and MCP-3/CCL7 at the protein (immunohistochemistry; n = 62) and transcriptional levels (RTqPCR; n = 173) in colorectal polyps with reference to the polyp malignancy potential. All chemokines were significantly upregulated in polyps at the protein level but downregulated at the transcriptional level by 1.4-(CCL2), 1.7-(CCL7), and 2.3-fold (CCL8). There was an inverse relation between the immunoreactivity toward chemokine proteins and the number of corresponding transcripts in polyps (CCL2 and CCL7) or in normal mucosa (CCL8). The downregulation of chemokine transcripts correlated with the presence of multiple polyps (CCL2 and CCL8), a larger polyp size (CCL2, CCL7, and CCL8), predominant villous growth patterns (CCL2, CCL7 and CCL8), and high-grade dysplasia (CCL2 and CCL8). In conclusion, MCP-1/CCL2, MCP-2/CCL8, and MCP-3/CCL7 chemokines are counter-regulated at the protein and transcriptional levels. Chemokine-directed chemopreventive strategies should therefore directly neutralize MCP proteins or target molecular pathways contributing to their enhanced translation or reduced degradation, rather than aiming at CCL2, CCL7 or CCL8 expression.

Heat Shock Proteins HSPA1 and HSP90AA1 Are Upregulated in Colorectal Polyps and Can Be Targeted in Cancer Cells by Anti-Inflammatory Oxicams with Arylpiperazine Pharmacophore and Benzoyl Moiety Substitutions at Thiazine Ring.

Heat shock proteins HSPA1/Hsp70α and HSP90AA1/Hsp90α are crucial for cancer growth but their expression pattern in colorectal polyps or whether they can be modulated by oxicams is unknown. We quantified (RTqPCR) HSPA1 and HSP90AA1 expression in 50 polyp-normal pairs in relation to polyp malignancy potential and examined the effect of piroxicam, meloxicam and five novel analogues on HSPA1 and HSP90AA1 expression (mRNA/protein) in colorectal adenocarcinoma lines. HSPA1 and HSP90AA1 were upregulated in polyps by 3- and 2.9-fold. Expression ratios were higher in polyps with higher dysplasia grade and dominant villous growth pattern, mostly a result of diminished gene expression in normal tissue. Classic oxicams had negligible/non-significant effect on HSP expression. Their most effective analogue inhibited HSPA1 protein and gene by 2.5-fold and 5.7-fold in Caco-2 and by 11.5-fold and 6.8-fold in HCT116 and HSPA1 protein in HT-29 by 1.9-fold. It downregulated HSP90AA1 protein and gene by 1.9-fold and 3.7-fold in Caco-2 and by 2-fold and 5.0-fold in HCT116. HSPA1 and HSP90AA1 are upregulated in colorectal polyps reflecting their potential for malignancy. HSPA1 in cancer cells and, to lesser degree, HSP90AA1 can be reduced by oxicam analogues with thiazine ring substituted via propylene linker by arylpiperazine pharmacophore with fluorine substituents and by benzoyl moiety.

S1493 A Benign Common Bile Duct Tumor: A Diagnostic Dilemma

Introduction: Benign extrahepatic bile duct tumors are uncommon, accounting for 6% of all extrahepatic bile duct masses. They tend to occur later in life and can present early with obstructive jaundice. They are morphologically and histologically similar with each other which makes it challenging to make an accurate diagnosis. These lesions can progress to malignancy so timely diagnosis and appropriate treatment is critical. We present a case of a benign common bile duct (CBD) tumor presenting in a healthy young man that had both features of a papillary and a tubulo-villous neoplasia, causing a diagnostic dilemma. Case Description/Methods: Patient is a 48-year-old man with history of pancreatic DB-IPMN, presenting with a one-month history of post-prandial upper abdominal pain. Physical exam was unremarkable. Labs showed elevated liver enzymes with ALT, AST, and ALP of 186, 78 U/L, 537 U/L, respectively; total bilirubin was 1 mg/dL. Abdominal MRI showed diffuse dilation of intrahepatic and extrahepatic ducts, as well as dilation of the pancreas duct with a prominent ampulla. ERCP showed a bulging ampulla with abnormal appearing tissue in the distal CBD [Figure 1]. Biopsy of the of distal CBD revealed intra-papillary neoplasm of bile duct (IPNB) without features of high-grade dysplasia [Figure 2A]. A fully covered metal stent was placed with immediate improvement in symptoms and laboratory markers. Case reviewed in tumor board which recommended obtaining more tissue prior to definitive management. A repeat ERCP with SpyGlass cholangioscopy-directed biopsies and biopsies of the ampulla revealed tubule-villous adenoma with high grade dysplasia [Figure 2B]. The patient underwent curative pancreaticoduodenectomy. Final histology did not reveal invasive cancer. Discussion: IPNB is an uncommon benign extra-hepatic bile duct tumor, characterized by papillary or villous growth within the bile duct lumen. It is more common in elderly men and can progress from atypia to invasive cholangiocarcinoma. About half of IPNBs show stromal invasion at the time of surgical resection. Accurate and timely diagnosis before metastasis is critical as surgical resection has excellent outcome with 5-year tumor free survival being 80%. Our case highlights an IPNB of the distal CBD extending into the ampulla, masquerading as a more common adenoma of the duodenal ampulla.Figure 1.: ERCP showing prominent ampulla (1a) and biliary sphincterotomy (1b)Figure 2.: Figure 2A: (Biospy#1) Intraductal papillary Neoplasia (papillary proliferation of biliary epithelium along with delicate fibrovascular cores) Figure 2B: (Biospy#2) (a) At low power (40x) demonstrating tubulovillous adenoma with papillary features. Figure 2B: (Biospy#2) (b) At high power (400x) showing an area with high grade dysplasia with cribriforming, nuclear atypia, and abundant mitotic figures.

Biphasic Expression of Atypical Chemokine Receptor (ACKR) 2 and ACKR4 in Colorectal Neoplasms in Association with Histopathological Findings.

Facilitating resolution of inflammation using atypical chemokine receptors (ACKR) as an anticancer strategy is considered but requires a deeper understanding of receptor role in carcinogenesis. We aimed at transcriptional analysis (RTqPCR) of ACKR2 and ACKR4 expression in colorectal adenoma-adenocarcinoma sequence in paired normal-neoplastic tissues from 96 polyps and 51 cancers. On average, ACKR2 was downregulated in neoplastic as compared to non-affected tissue in polyp (by 2.7-fold) and cancer (by 3.1-fold) patients. The maximal downregulation (by 8.2-fold) was observed in adenomas with the highest potential for malignancy and was gradually lessening through cancer stages I-IV, owing to increased receptor expression in tumors. On average, ACKR4 was significantly downregulated solely in adenocarcinomas (by 1.5-fold), less so in patients with lymph node metastasis, owing to a gradual decrease in ACKR4 expression among N0-N1-N2 cancers in non-affected tissue without changes in tumors. In adenomas, ACKR4 downregulation in neoplastic tissue increased with increasing potential for malignancy and contribution of villous growth pattern. ACKR4 expression increased in non-affected tissue with a concomitant decrease in pathological mucosa. In conclusion, the changes in ACKRs expression occur already in precancerous colorectal lesions, culminating in the adenomas with the highest potential for malignancy. Therefore, chemoprevention by manipulating ACKRs’ expression is worth exploration.

Clinical characteristics and long-term health in celiac disease patients diagnosed in early childhood: Large cohort study

BackgroundEarly detection of celiac disease could theoretically prevent most of the disease-associated complications, but long-term effects of this approach are unclear. AimsTo investigate features at diagnosis and adulthood health in celiac disease patients diagnosed in early childhood in 1965–2014. MethodsMedical data on 978 pediatric patients were collected and study questionnaires sent to 559 adult patients who were diagnosed in childhood. Results were compared between patients diagnosed in early (≤3.0 years) and later (3.1-17.9 years) childhood. ResultsEarly diagnosed patients (n=131) had more often total villous atrophy (37% vs 25%, p=0.001), gastrointestinal presentation (61% vs 47%, p<0.001), growth disturbances (70% vs 32%, p=0.001) and severe symptoms (30% vs 9%, p<0.001) and were less often screen-detected (10% vs 27%, p<0.001) at diagnosis than those diagnosed later (n=847).Among 239 adult responders, early diagnosed patients (n=36) had fewer comorbidities (33% vs 53%, p=0.034) but considered their health less often good/excellent (69% vs 84%, p=0.029). The groups were comparable in current age, dietary adherence, symptoms and health-related quality of life. ConclusionDespite more severe initial presentation, the long-term health in early diagnosed patients was mostly comparable or even better to those diagnosed later in childhood. Poorer self-perceived health suggests a need for support during the transition to adulthood care.

How does intestinal-type intraductal papillary mucinous neoplasm emerge? CDX2 plays a critical role in the process of intestinal differentiation and progression.

Intestinal-type intraductal papillary mucinous neoplasm (IPMN) of the pancreas is clinicopathologically distinctive. Our research aimed to elucidate the molecular mechanism of the development and progression of the intestinal-type IPMN. In 60 intestinal-type IPMN specimens, histological transitions from gastric-type epithelia to intestinal-type epithelia were observed in 48 cases (80%). CDX2/MUC2/alcian blue triple staining indicated that CDX2 appeared to precede MUC2 expression and subsequent alcian blue-positive mucin production. Expression of p21 and Ki-67 seemed to be accelerated by CDX2 expression (p = 6.02e-13 and p = 3.1e-09, respectively). p21/Ki-67 double staining revealed that p21 was mostly expressed in differentiated cells in the apex of papillae, while Ki-67 was expressed in proliferative cells in the base of papillae. This clear cellular arrangement seemed to break down with the progression of atypical grade and development of invasion (p = 0.00197). Intestinal-type IPMNs harbored frequent GNAS mutations (100%, 25/25) and RNF43 mutations (57%, 8/14) and shared identical GNAS and KRAS mutations with concurrent gastric-type IPMNs or incipient gastric-type neoplasia (100%, 25/25). RNF43 mutations showed emerging or being selected in intestinal-type neoplasms along with ß-catenin aberration. Activation of protein kinase A and extracellular-regulated kinase was observed in CDX2-positive intestinal-type neoplasm. These results suggest that gastric-type epithelia that acquire GNAS mutations together with induction of intrinsic CDX2 expression may evolve with clonal selection and additional molecular aberrations including RNF43 and ß-catenin into intestinal-type IPMNs, which may further progress with complex villous growth due to disoriented cell cycle regulation, acceleration of atypical grade, and advance to show an invasive phenotype.

The relationship between villous height and growth performance, small intestinal mucosal enzymes activities and nutrient transporters expression in weaned piglets.

Intestinal villi are important structures for digesting and absorbing nutrients. It was hypothesized that intestinal villous height (VH) is related to growth performance, and that VH is associated with digestive and absorptive capabilities in piglets. Sixty 21-d-old weaned piglets were fed identical diets for 28days. The piglets were grouped according to jejunal or ileal VH at the end of the experiment, including short, middle and high VH groups. The final BW (quadratic, p=.003), ADG (liner, p=.052; quadratic, p=.030) and G: F (liner, p=.074; quadratic, p=.005) were greater in higher VH piglets compared with the shorter VH piglets. Significant linear relationships were recorded between jejunal VH and maltase (linear, p=.003) and sucrase (linear, p=.004) activities. Besides, jejunal VH was significantly associated with maltase (r=0.357, p=.007), sucrase (r=0.394, p=.003) and alkaline phosphate (r=0.288, p=.033) activities. Jejunal Slc1a1 (linear, p=.004) mRNA expression increased as VH increased, while Slc7a1 (linear, p=.007) expression decreased as VH increased, and Slc1a1 (r=0.292, p=.031) expression was positively related to VH. In the ileum, Slc5a1 (linear, p=.002) and Slc1a1 (linear, p<.001) expression increased as VH increased, whereas Slc7a1 (linear, p=.045) expression decreased as VH increased. A positive relationship between ileal VH and Slc5a1 (r=0.331, p=.018), Slc1a1 (r=0.444, p=.001) and Slc6a19 (r=0.314, p=.026), respectively, was established. Additionally, Slc7a9 (r=0.0271, p=.057) expression tended to be positively associated with VH in ileum. In conclusion, the growth performance of piglets increased with increasing VH, which may via affecting mucosal enzymes activities and nutrient transporters' mRNA expression.

Colorectal Cancer Incidence and Mortality After Removal of Adenomas During Screening Colonoscopies

Recommendation of surveillance colonoscopy should be based on risk of colorectal cancer and death after adenoma removal. We aimed to develop a risk classification system based on colorectal cancer incidence and mortality following adenoma removal. We performed a multicenter population-based cohort study of 236,089 individuals (median patient age, 56 years; 37.8% male) undergoing screening colonoscopies with adequate bowel cleansing and cecum intubation at 132 centers in the Polish National Colorectal Cancer Screening Program, from 2000 through 2011. Subjects were followed for a median 7.1 years and information was collected on colorectal cancer development and death. We used recursive partitioning and multivariable Cox models to identify associations between colorectal cancer risk and patient and adenoma characteristics (diameter, growth pattern, grade of dysplasia, and number of adenomas). We developed a risk classification system based on standardized incidence ratios, using data from the Polish population for comparison. The primary endpoints were colorectal cancer incidence and colorectal cancer death. We identified 130 colorectal cancers in individuals who had adenomas removed at screening (46.5 per 100,000 person-years) vs 309 in individuals without adenomas (22.2 per 100,000 person-years). Compared with individuals without adenomas, adenomas ≥20 mm in diameter and high-grade dysplasia were associated with increased risk of colorectal cancer (adjusted hazard ratios 9.25; 95% confidence interval [CI] 6.39-13.39, and 3.58; 95% CI 1.96-6.54, respectively). Compared with the general population, colorectal cancer risk was higher or comparable only for individuals with adenomas ≥20 mm in diameter (standardized incidence ratio [SIR] 2.07; 95% CI 1.40-2.93) or with high-grade dysplasia (SIR 0.79; 95% CI 0.39-1.41), whereas for individuals with other adenoma characteristics the risk was lower (SIR 0.35; 95% CI 0.28-0.44). We developed a high-risk classification based on adenoma size ≥20 mm or high-grade dysplasia (instead of the current high-risk classification cutoff of ≥3 adenomas or any adenoma with villous growth pattern, high-grade dysplasia, or ≥10 mm in diameter). Our classification system would reduce the number of individuals classified as high-risk and requiring intensive surveillance from 15,242 (36.5%) to 3980 (9.5%), without increasing risk of colorectal cancer in patients with adenomas (risk difference per 100,000 person-years, 5.6; 95% CI -10.7 to 22.0). Using data from the Polish National Colorectal Cancer Screening Program, we developed a risk classification system that would reduce the number of individuals classified as high risk and require intensive surveillance more than 3-fold, without increasing risk of colorectal cancer in patients with adenomas. This system could optimize the use of surveillance colonoscopy.

Abstract 4213: Stool-derived eukaryotic RNA (seRNA) assay for noninvasive detection of colorectal cancer and high-risk adenomas

Abstract Colorectal cancer (CRC) is the second leading cause of cancer related deaths in the United States. The high mortality rate for CRC is largely attributable to the frequency of late-stage diagnoses, caused by low patient compliance with screening guidelines. A novel nucleic acid extraction method that isolates stool-derived eukaryotic RNA (seRNA) has permitted development of a colorectal cancer stool diagnostic test (CRC-SDT) that uses a fecal immunochemical test (FIT) and 11 seRNA transcripts to serve as a reliable and noninvasive screening alternative to lower mortality associated with CRC. Stool samples were obtained from 190 individuals prior to undergoing a screening colonoscopy. Patients were diagnosed as either having colorectal cancer, having high-risk adenomas (high-grade dysplasia, villous growth pattern, or &amp;gt;1.0cm in size), or healthy (low-risk adenomas, benign polyps, and/or no findings on a colonoscopy). FITs were obtained for each sample. Samples that had a positive FIT were considered positive for the CRC-SDT. Samples that had a negative FIT underwent seRNA isolation, library preparation (Illumina TruSeq Targeted RNA) and subsequent sequencing (Illumina NextSeq 550). A random forest model was built using 11 transcripts that were differentially expressed (log2 fold-change &amp;gt; 1; ANOVA p &amp;lt; 0.05) between diseased patients (CRC and high-risk adenomas) relative to the healthy cohort. Internal 10-fold cross-validation was performed for the training set and a receiver operating characteristic (ROC) area under the curve (AUC) determined model accuracy for cancer and high-risk adenomas relative to the healthy cohort. Model predictions for an independent hold-out test set were used to determine model accuracy for colorectal cancer and high-risk adenomas. Internal cross-validation for 154 samples attained a ROC AUC of 0.82. The trained model was employed on the independent hold-out test set of 36 samples and model predictions were compared to colonoscopy results. The model attained a 100% sensitivity for CRC (n = 7), a 60% sensitivity for high-risk adenomas (n = 5), and an 88% combined specificity for the healthy cohort (n = 24). CRC-SDT uses 12 biomarkers (FIT and expression from 11 seRNA transcripts) to accurately detect CRC and high-risk adenomas. Use of seRNA biomarkers dramatically improves detection of high-risk adenomas relative to existing noninvasive screening methods for CRC. Citation Format: Erica K. Barnell, Yiming Kang, Katie M. Campbell, Kimberly R. Kruse, Andrew R. Barnell, Elizabeth M. Wurtzler. Stool-derived eukaryotic RNA (seRNA) assay for noninvasive detection of colorectal cancer and high-risk adenomas [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 4213.

Characterization of Intraductal Papillary Neoplasm of the Bile Duct with Respect to the Histopathologic Similarities to Pancreatic Intraductal Papillary Mucinous Neoplasm.

Intraductal papillary neoplasms of the bile duct (IPNBs) are known to show various pathologic features and biological behaviors. Recently, two categories of IPNBs have been proposed based on their histologic similarities to pancreatic intraductal papillary mucinous neoplasms (IPMNs): type 1 IPNBs, which share many features with IPMNs; and type 2 IPNBs, which are variably different from IPMNs. The four IPNB subtypes were re-evaluated with respect to these two categories. Intestinal IPNBs showing a predominantly villous growth may correspond to type 1, while those showing papillay-tubular or papillay-villous growth correspond to type 2. Regarding gastric IPNB, those with regular foveolar structures with varying numbers of pyloric glands may correspond to type 1, while those with papillary-foveolar structures with gastric immunophenotypes and complicated structures may correspond to type 2. Pancreatobiliary IPNBs that show fine ramifying branching may be categorized as type 1, while others containing many complicated structures may be categorized as type 2. Oncocytic type, which displays solid growth or irregular papillary structures, may correspond to type 2, while papillary configurations with pseudostratified oncocytic lining cells correspond to type 1. Generally, type 1 IPNBs of any subtype develop in the intrahepatic bile ducts, while type 2 IPNBs develop in the extrahepatic bile duct. These findings suggest that IPNBs arising in the intrahepatic ducts are biliary counterparts of IPMNs, while those arising in the extrahepatic ducts display differences from prototypical IPMNs. The recognition of these two categories of IPNBs with reference to IPMNs and their anatomical location along the biliary tree may deepen our understanding of IPNBs.

Elevated systemic interleukin-7 in patients with colorectal cancer and individuals at high risk of cancer: association with lymph node involvement and tumor location in the right colon

Interleukin (IL)-7 is a cytokine essential for protective immunity, and it is considered as a promising agent for cancer immunotherapy. Recent studies, however, appear to associate IL-7 with aggressiveness of solid tumors. The IL-7 has been less studied in colorectal cancer (CRC) and conditions associated with increased risk of CRC development. To explore IL-7 status in bowel diseases, it was measured immunofluorometrically in 431 individuals (110 with CRC) by using Luminex platform. A level of IL-7 in CRC patients was significantly higher than in controls, did not differ from those with adenomas, but was lower than in both active and inactive inflammatory bowel disease (IBD) cases. In CRC, IL-7 was higher in patients with lymph node and distant metastases and with tumors located in right colon. In adenomas, IL-7 elevation was associated exclusively with villous growth pattern, while in IBD, circulating IL-7 reflected clinical activity of Crohn’s disease and ulcerative colitis. Systemic TNFα, IL-10, and PDGF-BB were independent predictors of circulating IL-7. In summary, our study is the first to demonstrate IL-7 elevation in CRC in association with metastatic disease and tumor location. Both associations should be considered when designing IL-7-based immunotherapies for CRC. Further studies on IL-7 functionality in CRC are necessary.

The Long and Short of It: A Case of Long-term Teduglutide for the Treatment of Intestinal Rejection in Small Bowel Transplant: 2016 ACG Presidential Poster Award

Small bowel transplantation (SBT) is a therapeutic option for those patients who suffer from short bowel syndrome, intestinal failure, and life-threatening complications related to total parental nutrition (TPN). Post-small bowel transplant rejection continues to be a major cause of morbidity and mortality. Endoscopically, graft rejection is characterized by hemorrhage, blunting, and sloughing of the intestinal villi while histologically, it is characterized by increased apoptotic bodies and decreased mitotic bodies per crypt. Teduglutide, a glucagon-like peptide 2 analogue, has been shown to improve intestinal absorption by promoting villous hypertrophy and crypt proliferation. We hypothesized that teduglutide 5 mg subcutaneously daily can promote small bowel villous growth and attenuate small bowel transplant rejection by increasing crypt division while decreasing the apoptotic rate. This novel application of teduglutide may facilitate recovery from rejection and improve nutritional absorption. We present an interesting case of a 22-year-old woman with a myofibroblastic tumor of the mesentery who underwent extensive small bowel resection and subsequent living related small bowel transplantation. She presented with fever and abdominal pain post-operative day 14 after SBT. She underwent ileoscopy revealing ulcerated and extensively denuded intestinal mucosal villi. Histology demonstrated inflammatory changes extending through the muscularis mucosa that was consistent with severe acute graft rejection. She was subsequently placed on teduglutide while continuing immunosuppressive therapy and underwent serial endoscopic evaluations with random small bowel biopsies. Endoscopically, there was serial improvement in the health of the small bowel graft, as histologically demonstrated by an increase in villous growth, overall decrease in the apoptotic rate, and overall increase mitotic rate over 9 months. Furthermore, we conducted novel fluorescent in-situ hybridization (FISH) analysis which confirmed an XY karyotype in the regenerating stem cells of the small bowel graft, indicating that her father's stem cells from the donated small bowel were indeed regenerating. In conclusion, we present a novel application of teduglutide to mitigate small bowel transplant rejection and promote villous growth. Teduglutide should be considered in these patients to help improve nutritional status, allow them to wean off TPN, and ultimately improve quality of life.

33 Placental maturation analysis in cases of term intra-uterine growth restriction revealed decreased villous angiogenesis and increased trophoblast turnover

Introduction Intra-uterine growth restriction (IUGR), failure of the fetus to reach normal birth weight, represents an important health hazard among neonates, and is associated with increased mortality and morbidity later in life. It is postulated that IUGR is associated with an inefficient blood and gas-supply at the feto-maternal interface of the placenta resulting in fetal hypoxia and under nutrition. Objectives The aim of the present study is to analyze, placental maturation assuring adequate increase of gas exchange efficiency by histo-morphological analyses of placentas comparing control term placentas to those of term growth restricted fetuses. This includes analyses of the degree of terminal villi vascularization, the diffusion distance between fetal capillary and maternal blood and the syncytial trophoblast turnover. Methods Paraffin sections of placental samples from term IUGR ( n = 10) and healthy term pregnancies ( n = 11) were stained immunohistochemically for endothelial cells (CD31). This is followed by stereological evaluation by light microscopy and an Image Analyzer. Surface areas, distances and numbers from terminal villi (TV), numbers and area of fetal capillaries (FC), diffusion distances (DD), vascular-syncytial membranes (VSM) and apoptotic knots (AK), were recorded. Non-parametric analyses were applied and statistical significance was set at P Results The mean birth weight for the IUGR group (2353±285g) in accordance to the group definitions differed significantly ( p p = 0.049). No significant differences were found in maternal age ( p = 0.28), BMI ( p = 0.24) gravidity or parity. We observed less TV per field ( p = 0.058), VSM’s ( p = 0.31) and FC’s per TV (0.64) less area of FC per TV ( p = 0.27) and shorter diffusion distances in IUGR compared to controls, however these differences are not statistically significant. VSM lengths were significantly shorter per TV ( p = 0.04) in IUGR. Analysis of AK showed a significant increase ( p = 0.002) in IUGR compared to controls. Conclusion TV’s are sites specialized in gas exchange. They comprise of VSM’s characterized by fusions of the basal membranes of the capillary endothelium and the syncytiotrophoblast layer making the diffusion distance shortest and thus gas exchange most efficient. There are less TV’s and shorter VSM’s in the IUGR samples. This diffusion deficit may be compensated, by decreasing the mean diffusion distance of all fetal capillaries. In addition the trophoblast turnover as measured by the presence of AK seems to be increased in IUGR. The preceding is supported by the theory of post-placental hypoxia. According to this theory both mother and placenta are normoxic whereas the fetus is hypoxic from fetoplacental malperfusion. This results in a reduced oxygen extraction from the placenta resulting in intra-placental hyperoxia. These high pO2 values inhibit villous growth resulting in decreased TV and shorter VSM’s, and accordingly cause IUGR. The high intra-placental pO2 leads to increased oxidative stress in the placenta and might also be the cause of the increased trophoblast turn over observed in IUGR.

Prokineticin1 and pregnancy

Prokineticin 1 (PROK1), also called EG-VEGF, is a peptide of 86 amino acids with multiple biological functions. PROK1 acts via two G-protein coupled receptors: PROKR1 PROKR2. PROK1 is highly expressed in the placenta. This article reports the expression and the role of PROK1 during normal and pathological pregnancies: (i) during early pregnancy, PROK1 exhibits a peak of placental expression shortly before the establishment of the feto-maternal circulation; (ii) its receptors, PROKR1 PROKR2 are highly expressed in human placenta; (iii) its expression is increased by hypoxia; (iv) PROK1 inhibits extravillous trophoblasts migration and invasion and increases their proliferation and survival; (v) PROK1 is also a pro-angiogenic placental factor that increases microvascular placental endothelial cells proliferation, migration, invasion, and permeability. Circulating PROK1 levels are five times higher in pregnant women during the first trimester compared to the second and third trimesters. Also, its serum levels are higher in patients with preeclampsia (PE) and in patients with isolated intra-uterine growth restriction (IUGR). In mice, maintaining high level of PROK1 beyond its normal period of production (>10.5dpc) reproduces symptoms of PE. To date, our results demonstrated that PROK1 is a central factor of human placentation with direct roles both in the control of trophoblast invasion and villous growth. Thus, a failure in the expression of PROK1 and/or its receptor during pregnancy may contribute to the development of PE and/or IUGR. Besides theses original findings, we also report a direct role of this factor in parturition.