Ductal Invasion Research Articles

New Method of Papillectomy May Decrease Recurrence: Anchoring Method versus Conventional Method.

Background/Objectives: Endoscopic papillectomy (EP) is the preferred treatment for ampullary tumors because it has fewer side effects than surgical removal. This study retrospectively compared a new anchoring EP method (A-EP) with the conventional (C-EP) approach. Methods: Ninety-nine patients who underwent EP at a single medical institution between 2009 and 2021 were retrospectively reviewed. In all patients, the indications for EP were pathological adenoma with <10 mm of biliary invasion and a tumor diameter <30 mm on endoscopic ultrasonography. The exclusion criteria were antiplatelet/anticoagulant use, previous upper GI surgery, or prior biliary/pancreatic endoscopic therapy. One expert endoscopist performed the two types of EPs, A-EP and C-EP. Results: Sixty-two patients underwent A-EP, and 37 underwent C-EP. There were no significant differences in baseline characteristics, such as sex, age, tumor size, and ductal invasion on endoscopic ultrasound. The A-EP group had higher en bloc resection rates (95.2% vs. 78.4%, p = 0.010). Although the difference was not statistically significant, it tended towards fewer incidences of pancreatitis (p = 0.081) and duct stricture (p = 0.081). The recurrence rate was lower in the A-EP group (8.1% vs. 37.8%, p = 0.000). There were no significant differences between the two groups regarding the follow-up period (A-EP vs. C-EP, 725 vs. 1045 days, p = 0.109) or the days of recurrence (A-EP vs. C-EP, 341 vs. 562 days, p = 0.551). Conclusions: A-EP showed better outcomes than C-EP in terms of en bloc resection and recurrence rates, providing evidence for the efficacy of this novel EP method.

Abstract 3463: Nischarin agonist rilmenidine inhibits pancreatic ductal adenocarcinoma cell migration and invasion

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive lethal malignancy due to the lack of early diagnosis and limited response to all treatment modalities. Early metastatic dissemination, rich desmoplastic stroma and immunosuppressive infiltrate are some of the features contributing to the poor prognosis of PDAC patients (5-year survival rate of less than 10%). Despite scientific progress in the development of new therapeutic regimes, PDAC patients in low- and middle-income countries have restricted access to high-cost innovative drugs. Recently, attention has been given to the concept of drug repurposing that could meet the need for novel but affordable and widely accessible therapies. Imidazoline receptor 1 Nischarin (NISCH, IR1, IRAS) is an integrin α5-interacting protein that acts as a tumor suppressor in breast cancer through modulation of cancer cell motility and survival. Importantly, NISCH has several clinically approved agonists that are used for the treatment of hypertension and are shown to modulate autophagy and reduce inflammation and fibrosis, which makes NISCH a potentially good therapeutic target for defying PDAC progression. Based on our GSEA analysis of NISCH expression in PDAC patient samples, NISCH was predicted to have an effect on cancer cell focal adhesion assembly and cytoskeletal organization. Therefore, we examined the effects of NISCH agonists on PDAC cells fitness in vitro. A panel of PDAC cell lines was tested in MTT assay to determine the effects of NISCH agonists rilmenidine, clonidine and moxonidine on cell viability. Flow cytometry, western blot and immunofluorescence were used to examine the effects of NISCH agonists on PDAC cell metabolism, migration and cytoskeletal organization. Tg(fli1:EGFP) zebrafish model was used to examine the effects of NISCH agonists on PDAC cell growth and invasion. We found that NISCH was expressed in PDAC cell lines, and that out of the three tested agonists, rilmenidine most potently inhibited cancer cell viability. Treatment with rilmenidine significantly reduced PANC-1, MIA PaCa-2 and BxPC-3 cell attachment to extracellular matrix (collagen type I and fibronectin) and migratory potential in wound healing and transwell assays. Furthermore, treatment with rilmenidine altered the organization of actin cytoskeleton and focal adhesion assembly, potentially through the reduction of intracellular reactive oxygen species (ROS) levels. Importantly, treatment with rilmenidine inhibited invasion of PANC-1 cells in Tg(fli1:EGFP) zebrafish model. With these antimigratory and potentially antimetastatic effects of rilmenidine, our study lays the ground for a more extensive examination of the biological role of NISCH in PDAC progression and implies that its agonists may be good candidates for drug repurposing in this type of cancer. Citation Format: Marijana Pavlovic, Marija Ostojic, Kristina Zivic, Aleksandar Pavic, Tatjana Srdic-Rajic, Jelena Grahovac. Nischarin agonist rilmenidine inhibits pancreatic ductal adenocarcinoma cell migration and invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3463.

Repression of endogenous retroviruses prevents antiviral immune response and is required for mammary gland development

The role of heterochromatin in cell fate specification during development is unclear. We demonstrate that loss of the lysine 9 of histone H3 (H3K9) methyltransferase G9a in the mammary epithelium results in de novo chromatin opening, aberrant formation of the mammary ductal tree, impaired stem cell potential, disrupted intraductal polarity, and loss of tissue function. G9a loss derepresses long terminal repeat (LTR) retroviral sequences (predominantly the ERVK family). Transcriptionally activated endogenous retroviruses generate double-stranded DNA (dsDNA) that triggers an antiviral innate immune response, and knockdown of the cytosolic dsDNA sensor Aim2 in G9a knockout (G9acKO) mammary epithelium rescues mammary ductal invasion. Mammary stem cell transplantation into immunocompromised or G9acKO-conditioned hosts shows partial dependence of the G9acKO mammary morphological defects on the inflammatory milieu of the host mammary fat pad. Thus, altering the chromatin accessibility of retroviral elements disrupts mammary gland development and stem cell activity through both cell-autonomous and non-autonomous mechanisms.

Adenomatous neoplasia in the papilla of Vater endoscopic and/or surgical resection?

Adenomatous neoplasia in the papilla of Vater needs to be resected in order not to progress. It can be challenging to distinguish between early ampullary malignant lesions and non-invasive adenomas, due to the overlap in symptoms and radiological findings. This retrospective study describes the different findings and treatment decisions taken prior to endoscopic and/or surgical resection of ampullary adenomatous lesions. Patients treated with endoscopic and/or surgical resection for suspected or verified ampullary adenomatous neoplasia, between January 2006 and July 2018, where pre-interventional cross-sectional imaging could not discern an obvious invasive, malignant tumor, were included. Findings were compared against the final diagnosis of the histopathological analysis on the resected specimen. In total, 172 met the inclusion criteria. Patients were treated with either surgical resection (n = 96), endoscopic papillectomy (EP) (n = 55) or both (n = 21). The final diagnosis was in 48% ampullary adenocarcinoma, and the remaining had either ampullary adenoma (38%) or non-neoplastic lesions (14%). In patients where symptoms and cross-sectional imaging were suspicious for malignancy, but with no tissue samples that confirmed neoplasia prior to surgical resection, only 47% had adenocarcinoma. The remaining had either adenoma (9%) or non-neoplastic lesions (44%). Adenocarcinoma was revealed in 27% of the patients where endoscopic biopsies had shown adenoma. Patients with adenoma, treated with EP, were cured in 59%. However, 28% were after EP sent for further surgery due to ductal invasion or a finding of adenocarcinoma. In patients with a suspicion of ampullary neoplasia on imaging, attempts should be made to get endoscopic tissue samples before deciding on a treatment strategy. If biopsies show ampullary adenoma, patients should be considered for EP, unless there are clear radiologic or endoscopic signs of malignancy. Patients with adenocarcinoma on endoscopic biopsies should undergo surgical resection.

Extraction of intra-biliary hepatocellular carcinoma by endoscopic retrograde cholangiopancreatography

BackgroundHepatocellular carcinoma with biliary ductal invasion is rare and associated with a significantly lower survival rate.Case presentationWe present an unusual case of a patient with hepatocellular carcinoma and biliary invasion, who had his diagnosis confirmed by histological analysis from tissue extracted by endoscopic retrograde cholangiopancreatography. An 87-year-old male presented with a 1-day history of right upper quadrant pain and jaundice. His past medical history included recurrent gallstone cholangitis and a previous cholecystectomy. An abdominal CT demonstrated a dilated intrahepatic biliary tree with left proximal intrahepatic hyperdensities, as well as a 3 cm hepatocellular carcinoma. He was initially suspected to have concurrent gallstone cholangitis and a newly diagnosed hepatocellular carcinoma. Endoscopic retrograde cholangiopancreatography and balloon trawling of the intraductal lesions extracted necrotic tumour-like tissue which was histologically consistent with hepatocellular carcinoma. The extraction of the intra-biliary portion of HCC resulted in complete resolution of his jaundice, enabling further treatment with nivolumab, which would not have been possible if the obstruction was not cleared. The patient is currently well and has completed his 6th cycle of nivolumab.ConclusionObstructive jaundice is an uncommon presentation for patients with HCC. it is key for clinicians to be aware of the possibility of intrabiliary invasion in order obtain an early diagnosis and to reduce any delay in treatment.

A Rare Cause of Obstructive Jaundice: Is Surgery the Only Option?

Question: A 71-old men presented with a 2-week history of painless jaundice. Physical examination showed icteric sclera and abdominal examination revealed slight deep tenderness in epigastrium. Laboratory tests showed the following results: aminotransferase (AST), 251 IU/L; alanine transaminase (ALT), 118 IU/L; serum total bilirubin, 4.0 mg/dL and carbohydrate antigen (CA) 19-9, 50.0 U/mL. Also, serum total IgG revealed 1380.90 mg/dL with 135.2mg/dL of IgG4 subclass. Abdominal computed tomography (CT) revealed a relatively well-defined mass with homogeneous density in pancreatic head causing bile and pancreatic duct dilatation (Figure A). The positron emission tomography (PET) CT revealed about 5cm focal abnormal accumulation of fluorodeoxyglucose (FDG) in pancreatic head portion and periportal lymph node (Figure B). Endoscopic ultrasonography (EUS) revealed a heterogeneous low echogenic mass in pancreatic head with multifocal hyperechoic foci and marked bile duct dilatation with ductal invasion (Figure C). EUS-guided tissue acquisition and endoscopic retrograde cholangio-pancreatography was performed. The cholangiography shows short segment stricture at distal common bile duct with upstream ductal dilatation (Figure D), subsequently, decompression with plastic stent placement for obstructive jaundice was done. Histologic findings from EUS-guided tissue acquisition and endo-biliary biopsy revealed abundant infiltration of IgG4 positive plasma cells and lymphocytes around the pancreatic duct with storiform fibrosis (Figure E and F). CME Exam 1: A Rare Cause of Obstructive Jaundice: Is Surgery the Only Option?GastroenterologyVol. 158Issue 8Preview Full-Text PDF

A Multiscale Agent-Based Model of Ductal Carcinoma In Situ.

Objective:we present a multiscale agent-based model of Ductal Carcinoma in Situ (DCIS) in order to gain a detailed understanding of the cell-scale population dynamics, phenotypic distributions, and the associated interplay of important molecular signaling pathways that are involved in DCIS ductal invasion into the duct cavity (a process we refer to as duct advance rate here).Methods:DCIS is modeled mathematically through a hybridized discrete cell-scale model and a continuum molecular scale model, which are explicitly linked through a bidirectional feedback mechanism.Results:we find that duct advance rates occur in two distinct phases, characterized by an early exponential population expansion, followed by a long-term steady linear phase of population expansion, a result that is consistent with other modeling work. We further found that the rates were influenced most strongly by endocrine and paracrine signaling intensity, as well as by the effects of cell density induced quiescence within the DCIS population.Conclusion:our model analysis identified a complex interplay between phenotypic diversity that may provide a tumor adaptation mechanism to overcome proliferation limiting conditions, allowing for dynamic shifts in phenotypic populations in response to variation in molecular signaling intensity. Further, sensitivity analysis determined DCIS axial advance rates and calcification rates were most sensitive to cell cycle time variation.Significance:this model may serve as a useful tool to study the cell-scale dynamics involved in DCIS initiation and intraductal invasion, and may provide insights into promising areas of future experimental research.

Effect of decreased activity of 5’-adenosine monophosphate activated protein kinase on breast development in adolescent mice

Objective To investigate the effects of decreased 5’-activated protein kinase (AMPK) activity in mouse mammary epithelium on the proliferation of ductal epithelial cells and the development of teminal end bud (TEB) in adolescent mice. Methods AMPK DN transgenic mice were successfully constructed. The fourth pair of mammary gland tissues from the right abdomen of 10 WT and DN mice were stained by whole tissue staining and PCNA immunohistochemical staining. The duct coverage area, initial duct length, number of ductal branches per millimeter, percentage of ductal invasion length, terminal tip of mammary gland tissues of WT and DN mice were analyzed. TEB average quantity, average maximum area of TEB. The ratio of PCNA positive cells to the total number of glandular epithelial cells was calculated. Results At 5th week, the number of ductal branches per millimeter in DN mice and WT mice was (1.39±0.18) mm vs. (1.21±0.11) mm, the average number of TEB was (13.99±3.14) vs. (10.65±2.09) and the average maximum area of TEB was (0.104±0.014) mm2 vs. (0.082±0.013) mm2, respectively, with significant difference (t=2.698, 2.800, 3.642, P<0.05); at 6th week, the number of ductal branches per millimeter in DN mice and WT mice was (1.59±0.013) mm2, respectively. 31) vs. (1.41±0.34) mm, the average number of TEB (15.86±3.26) vs. (12.47±2.34), the average maximum area of TEB (0.118±0.017) mm2vs. (0.087±0.014) mm2, the difference was statistically significant (t=2.406, 2.671, 4.451, P<0.05). The percentage of PCNA positive cells in mammary epithelial cells of 5 w DN mice (86.47±4.67) was significantly higher than that of WT mice (65.32±3.87)%, and 6 w DN mice (65.32±3.87)%. The percentage of cells in WT group was significantly higher than that in WT group (85.37±4.35) (63.47±3.45)%. The difference was statistically significant (t=4.614, 12.474, P< 0.05). Conclusion The decrease of AMPK activity in mouse mammary epithelium can promote the proliferation of ductal epithelial cells and the development of TEBs in adolescent mice. Key words: Mouse mammary epithelial cells; Adolescence; 5’-activated protein kinase; Proliferation; Teminal end bud

Sfrp3 modulates stromal\u2013epithelial crosstalk during mammary gland development by regulating Wnt levels

Mammary stroma is essential for epithelial morphogenesis and development. Indeed, postnatal mammary gland (MG) development is controlled locally by the repetitive and bi-directional cross-talk between the epithelial and the stromal compartment. However, the signalling pathways involved in stromal–epithelial communication are not entirely understood. Here, we identify Sfrp3 as a mediator of the stromal–epithelial communication that is required for normal mouse MG development. Using Drosophila wing imaginal disc, we demonstrate that Sfrp3 functions as an extracellular transporter of Wnts that facilitates their diffusion, and thus, their levels in the boundaries of different compartments. Indeed, loss of Sfrp3 in mice leads to an increase of ductal invasion and branching mirroring an early pregnancy state. Finally, we observe that loss of Sfrp3 predisposes for invasive breast cancer. Altogether, our study shows that Sfrp3 controls MG morphogenesis by modulating the stromal-epithelial cross-talk during pubertal development.

Hepatocelluar Carcinoma Invading the Gallbladder - A Rare Cause of Hemobilia

Bleeding from the hepatobiliary tract (hemobilia) is a rare cause of acute upper GI bleed. It should be considered in any patient with recent history of hepatic parenchymal or biliary intervention, angioembolization, blunt abdominal trauma or hepatic malignancy. We describe a rare case of hemobilia due to hepatocellular carcinoma (HCC) invading the gall bladder wall. A 52- year old female with Laennec's cirrhosis complicated by HCC status post transarterial chemoembolization (TACE) presented with black tarry stools, dizziness and abdominal pain. Physical examination was significant for right upper quadrant tenderness to palpation and melanotic stools. Labs were consistent with severe anemia, thrombocytopenia and liver profile derangement. An MRI performed the month prior to admission, demonstrated a 3.1 cm oval-shaped, arterial phase enhancing lesion in the lateral aspect of the right hepatic lobe, suspicious for residual HCC (Fig A). She was aggressively transfused multiple blood products, placed on a proton pump inhibitor and started on octreotide. An emergent EGD was performed which identified blood oozing from the ampulla of vater; consistent with a diagnosis of hemobilia (Fig B). A CTA was negative for active extravasation. The presumed etiology was invasion of the residual HCC into the gall bladder wall given the close proximity identified on MRI. Given her history of TACE and severe liver dysfunction, she was not a candidate for embolization and she was medically managed. First recorded in 1654 by Francis Glisson, hemobilia is an exceedingly rare condition with unknown exact incidence. Traumatic etiology accounts for 40-80 percent of all cases. In patients with HCC, invasion into the blood vessels is a frequent feature, however gallbladder and ductal invasion is widely unrecognized. One study involving 24 patients with HCC was performed to describe the features of intra-biliary duct invasion. 21 percent developed hemobilia; one patient subsequently died from hemorrhagic shock. Approximately one-third of all patients will present with quinke's Triad: acute upper GI bleed (100 percent), biliary colic (70 percent) and jaundice (60 percent). Diagnosis is by visualizing blood from the Ampulla of vater via EGD or ERCP. The best diagnostic and therapeutic intervention is angiography with embolization (success rate > 95 percent). Despite aggressive intervention, severe hemobilia has been shown to have a mortality as high as 25 percent.1950_A Figure 1. A 3.1 cm oval shaped arterial phase enhancing lesion in the lateral aspect of the right hepatic lobe (arrow) suspicious for residual HCC within close proximity of the gall bladder.1950_B Figure 2. EGD oozing from the ampulla of vater (A) and showing blood oozing within the second part of the duodenum (B).

Acute Upper Gastrointestinal Bleeding: Think Outside the Ampulla

Bleeding from the hepatobiliary tract (hemobilia), is a rare cause of an acute upper GI bleed. It should be considered in any patient with a recent history of hepatic parenchymal or biliary intervention, angioembolization, blunt abdominal trauma or hepatic malignancy. We describe a case of hemobilia due to hepatocellular carcinoma (HCC) invading the gallbladder wall.Figure. 3: 1 cm oval-shaped, arterial-phase enhancing lesion in the lateral aspect of the right hepatic lobe (arrow) suspicious for residual HCC within close proximity to the gallbladder.Figure: EGD demonstrating blood oozing within the 2nd portion on the duodenum (A) and oozing from the Ampulla of Vater (B).A 52 year-old female with PMHx of Laennec's cirrhosis complicated by HCC status post transarterial chemoembolization (TACE) presented with black tarry stools, dizziness and abdominal pain. Physical exam was significant for right upper quadrant tenderness to palpation and melanotic stools. Labs were consistent with severe anemia, thrombocytopenia and liver profile derangement. An MRI performed the month prior to admission, demonstrated a 3.1 cm oval-shaped, arterial-phase enhancing lesion in the lateral aspect of the right hepatic lobe, suspicious for residual HCC (fig. A). She was aggressively transfused multiple blood products and placed on a proton pump inhibitor and octreotide drip. An emergent EGD was performed which identified blood oozing from the Ampulla of Vader; consistent with a diagnosis of hemobilia (fig. B). A CTA was negative for active extravasation. The presumed etiology was invasion of the residual HCC into the gallbladder wall given the close proximity identified on MRI. Given her history of TACE and severe liver dysfunction, she was not a candidate for embolization and she was medically managed. First recorded in 1654 by Francis Glisson, hemobilia is an exceedingly rare condition with an unknown exact incidence. Traumatic etiology accounts for 40 - 80 percent of all cases. In patients with HCC, invasion into the blood vessels (most commonly portal vein) is a frequent feature, however gallbladder and ductal invasion is widely unrecognized. One study involving 24 patients with HCC was performed to describe the features of intra-bile duct invasion. 21% developed hemobilia; one patient subsequently died from hemorrhagic shock. Approximately one-third of all patients will present with Quincke's triad: acute upper GI bleed (100%), biliary colic (70%) and jaundice (60%). Diagnosis is by visualizing blood from the Ampulla of Vader via EGD or ERCP. The best diagnostic and therapeutic intervention is angiography with embolization (success rate > 95%). Despite aggressive intervention, severe hemobilia has been shown to have a mortality as high as 25%.

Ectodysplasin target gene Fgf20 regulates mammary bud growth and ductal invasion and branching during puberty

Mammary gland development begins with the appearance of epithelial placodes that invaginate, sprout, and branch to form small arborized trees by birth. The second phase of ductal growth and branching is driven by the highly invasive structures called terminal end buds (TEBs) that form at ductal tips at the onset of puberty. Ectodysplasin (Eda), a tumor necrosis factor-like ligand, is essential for the development of skin appendages including the breast. In mice, Eda regulates mammary placode formation and branching morphogenesis, but the underlying molecular mechanisms are poorly understood. Fibroblast growth factor (Fgf) receptors have a recognized role in mammary ductal development and stem cell maintenance, but the ligands involved are ill-defined. Here we report that Fgf20 is expressed in embryonic mammary glands and is regulated by the Eda pathway. Fgf20 deficiency does not impede mammary gland induction, but compromises mammary bud growth, as well as TEB formation, ductal outgrowth and branching during puberty. We further show that loss of Fgf20 delays formation of Eda-induced supernumerary mammary buds and normalizes the embryonic and postnatal hyperbranching phenotype of Eda overexpressing mice. These findings identify a hitherto unknown function for Fgf20 in mammary budding and branching morphogenesis.

Abstract 464: Novel role of xpo1 in regulating MicroRNAs related to pancreatic ductal adenocarcinoma invasion and metastasis

Abstract Objective: There are no known reports on the role of exportin 1 (XPO1; also known as chromosome maintenance region 1 [CRM1]) in microRNA biology. In this study, we for the first time demonstrate that interfering with XPO1 machinery can influence miRNA signaling leading to suppression of pancreatic ductal adenocarcinoma (PDAC) proliferation, invasion and metastasis. Methods: miRNA arrays (LCSciences, Houston, TX) were performed on total RNA samples from PDAC cell lines (HPAC, MiaPaCa-2, AsPc-1 and L3.6pl) and normal human pancreatic ductal epithelial (HPDE) cells. PDAC cells were treated with XPO1 inhibitor (Selinexor) or transfected with control siRNA, XPO1 siRNA (Santa Cruz), miR-control or miR-145 mimic (Applied biosystems) all at a final concentration of 20 nM using DharmaFact Transfection Reagent (Dharmacon, Lafayette. CO). The total RNA and total protein from treated or transfected cells were subjected to real-time PCR or immunoblot analysis in order to measure expression level of miR-145, let-7d, miR-34c, miR-320, miR-205, and miR-145 target or downstream genes including EGFR, MMP1, MT-MMP, c-Myc, Sox-2, and PAK4. The impact of XPO1 inhibitor Selinexor on PDAC growth, proliferation, invasion and migrations was also evaluated using MTT and scratch assay. Results: In this study, we show that PDAC cells have significantly reduced expression of miR-145 when compared to normal pancreatic duct epithelial cells. Similarly, forced expression of miR-145 in PDAC cells inhibited cell proliferation and migration. Conversely, we demonstrate that RNAi of XPO1 by siRNA knockdown or chemical inhibition of XPO1 by selective inhibitor of nuclear export compound (Selinexor) restores miR-145 expression in PDAC cells ultimately leading to inhibition of cell proliferation and migration. In addition, we show that the inhibition of cell proliferation and migration by Selinexor is mediated through the down-regulation of known miR-145 signals including EGFR, MMP1, MT-MMP, c-Myc, PAK4 and Sox-2. Selinexor also induced the expression of two important tumor suppressive miRNAs, miR-34c and let-7d, leading to the up-regulation of p21WAF1. We also observed the down-regulation of oncomir mir-205. Conclusions: These results are the first to show that targeted inhibition of the nuclear exporter protein XPO1 by RNAi or Selinexor could restore tumor suppressive miRs in PDAC. Citation Format: Asfar S. Azmi, Yiwei Li, Irfana Muqbil, Amro Aboukameel, William Senapedis, Erkan Baloglu, Yosef Landesman, Michael Kauffman, Sharon Shacham, Philip A. Philip, Ramzi M. Mohammad. Novel role of xpo1 in regulating MicroRNAs related to pancreatic ductal adenocarcinoma invasion and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 464. doi:10.1158/1538-7445.AM2017-464

Development of a three dimensional, multiscale agent-based model of ductal carcinoma in situ.

Ductal carcinoma in situ (DCIS) is the most commonly diagnosed form of non-invasive breast cancer, constituting 20% of all new breast cancer cases in the United States. Although non-invasive, DCIS is usually treated surgically through resection. Interestingly, long-term survival studies have shown that patient survival rates are not significantly impacted by the type or resection, indicating that increased breast conservation through minimized surgical resection may indeed be possible. This requires a greater understanding of disease development, so that clinicians may more accurately determine surgical margins which minimize patient impact while maintaining survival rates. To this end, we have developed a three-dimensional, lattice-free multiscale agent based model of DCIS designed to help quantify ductal invasion rates, in order to allow clinicians to better estimate disease age and extent of invasion, and to predict surgical margins based on parameters obtainable from non-invasive testing (i.e., mammography). Here, we present the model development to date, and discuss some preliminary results.

Role of peptidylarginine deiminase 2 (PAD2) in mammary carcinoma cell migration

BackgroundPenetration of the mammary gland basement membrane by cancer cells is a crucial first step in tumor invasion. Using a mouse model of ductal carcinoma in situ, we previously found that inhibition of peptidylarginine deiminase 2 (PAD2, aka PADI2) activity appears to maintain basement membrane integrity in xenograft tumors. The goal of this investigation was to gain insight into the mechanisms by which PAD2 mediates this process.MethodsFor our study, we modulated PAD2 activity in mammary ductal carcinoma cells by lentiviral shRNA-mediated depletion, lentiviral-mediated PAD2 overexpression, or PAD inhibition and explored the effects of these treatments on changes in cell migration and cell morphology. We also used these PAD2-modulated cells to test whether PAD2 may be required for EGF-induced cell migration. To determine how PAD2 might promote tumor cell migration in vivo, we tested the effects of PAD2 inhibition on the expression of several cell migration mediators in MCF10DCIS.com xenograft tumors. In addition, we tested the effect of PAD2 inhibition on EGF-induced ductal invasion and elongation in primary mouse mammary organoids. Lastly, using a transgenic mouse model, we investigated the effects of PAD2 overexpression on mammary gland development.ResultsOur results indicate that PAD2 depletion or inhibition suppresses cell migration and alters the morphology of MCF10DCIS.com cells. In addition, we found that PAD2 depletion suppresses the expression of the cytoskeletal regulatory proteins RhoA, Rac1, and Cdc42 and also promotes a mesenchymal to epithelial-like transition in tumor cells with an associated increase in the cell adhesion marker, E-cadherin. Our mammary gland organoid study found that inhibition of PAD2 activity suppresses EGF-induced ductal invasion. In vivo, we found that PAD2 overexpression causes hyperbranching in the developing mammary gland.ConclusionTogether, these results suggest that PAD2 plays a critical role in breast cancer cell migration. Our findings that EGF treatment increases protein citrullination and that PAD2 inhibition blocks EGF-induced cell migration suggest that PAD2 likely functions within the EGF signaling pathway to mediate cell migration.

Intraductal neoplasms of the bile duct. A new challenge to biliary tract tumor pathology.

Invasive biliary tract carcinomas are usually tubular adenocaricnomas with abundant desmoplastic reactions and frequent ductal and periductal invasion at the time of the diagnosis. Recently, several intraductal neoplasms of the bile duct, particularly at a pre-invasive stage, have been recognized. They include intraductal papillary neoplasm of the bile duct (IPNB), biliary intraepithelial neoplasm (BilIN), and others, such as intraductal tubulopapillary neoplasm (ITPN) of the bile duct. IPNBs are grossly visible predominantly intraductal-growing papillary neoplasms covered by well-differentiated neoplastic epithelium with fine fibrovascular cores in the dilated bile ducts. Regarding their similarities to intraductal papillary mucinous neoplasm of the pancreas (IPMN) of main pancreatic duct type, some IPNBs resemble IPMN ("pancreatic type"), while others are only somewhat similar or variably different from IPMN ("non-pancreatic type"). Some IPNBs develop via a common oncogenic signaling pathway, and others, particularly those of intestinal type, frequently show GNAS mutations, as in IPMN. BilINs are a microscopically recognizable flat or micropapillary pre-invasive neoplasm and are presumed to precede conventional nodular-sclerosing cholangiocarcinomas. ITPN of the bile duct is a rare neoplasm composed of densely packed tubular glands. These three types of neoplasms are not infrequently associated with invasive adenocarcinoma. Pre-invasive intraglandular neoplasms of the peribiliary glands, another epithelial system in the biliary tree, have been also reported. Further characterization of these intraductal and intraglandular neoplasms of the bile duct is needed to overcome devastating invasive biliary tract carcinoma.

Endoscopic Features and Management Strategies of Ampullary Carcinoids: A Case Series

Introduction: Although the gastrointestinal tract is the most common site for occurrence of carcinoids, particularly the small bowel and the appendix, ampullary carcinoids are rather rare, with less than 100 cases reported in literature, most of which are single case reports. We present our experience of managing three such cases, two by endoscopic ampullectomy and one requiring surgery. Case Presentation: The three patients presented with obstructive jaundice. Abdominal imaging suggested ampullary masses of varying sizes, between 1.2 to 1.8 cm, accompanied by both intra and extrahepatic biliary duct dilatation and pancreatic duct dilatation. On endoscopy the tumors were noted to be umbilicated and firm. On endoscopic ultrasound (EUS) the masses were hypoechoic and confined to the deep mucosa. No CBD involvement or periduodenal lymphadenopathy was noted in any of these cases. One patient elected for surgical treatment (Whipple's) and the tumor was successfully excised. The other two underwent successful endoscopic ampullectomy. The cut surface of the endoscopically resected lesions appeared yellow and granular. The tumor was confirmed on pathology to be well-differentiated low-grade carcinoid, confined to the deep mucosa. Both patients had an uneventful post-ampullectomy course with resolution of their jaundice. Surveillance endoscopy and biopsy showed no residual tumor. Discussion: Ampullary carcinoids are rare tumors, which were traditionally managed with pancreatoduodenectomy or local excision. EUS has changed this paradigm and gives discrete information including depth of tumor, ductal invasion, and lymph node involvement. Ampullary carcinoids can be identified endoscopically by the typical features described above and successfully managed by endoscopic ampullectomy.

Abstract 1627: Kinase-independent function of focal adhesion kinase in lung metastasis of breast cancer

Abstract Focal adhesion kinase (FAK) has been shown to promote mammary tumorigenesis, lung metastasis and the maintenance of cancer stem cells in the MMTV-PyMT breast cancer model. Using mammary epithelial cell (MaEC)-specific FAK knockout and kinase-defective (KD) knockin mouse models, we recently demonstrated that loss of FAK kinase activity in MaECs specifically impaired luminal progenitor proliferation and alveologenesis, whereas FAK kinase-independent function was sufficient to support ductal invasion and basal mammary stem cell activity. To further elucidate this kinase-independent role of FAK in mammary tumorigenesis and lung metastasis, we established and analyzed three cohorts of PyMT mice expressing wild type FAK (Ctl-MT), KD-FAK (cKD-MT) and FAK-null (cKO-MT) in MaECs. This study revealed that both FAK deletion and KD knockin mutation in MaECs significantly suppressed mammary tumorigenesis and the content of stem-like tumor cells (CD24+CD29+CD61+). However, in contrast to cKO-MT mice that displayed diminished metastatic nodule formation in the lungs, loss of FAK kinase activity by KD knockin mutation did not affect lung metastatic nodule formation. Using a Boyden chamber assay, we further demonstrated that, unlike cKO-MT tumor cells that displayed diminished cell migration induced by fibronectin or TGFβ1 (as compared to Ctl-MT tumor cells), cKD-MT tumor cells exhibited strong fibronectin-, but not TGFβ1-induced migratory activity. Immunofluorescence of primary tumor sections also revealed that both Ctl-MT and cKD-MT tumors displayed strong ERK activation at tumor periphery, whereas cKO-MT tumors had weak ERK activation in this region. Lastly, when treated with or without paclitaxel in culture, we found that paclitaxel was able to induce strong expression of cleaved caspase 3 and p21 in cKO-MT but not Ctl-MT and cKD-MT tumor cells, suggesting a role for FAK kinase-independent function in the suppression of p21 expression and protection of apoptosis. Together, this study strongly implicates a role for FAK kinase-independent function in promoting lung metastasis. Citation Format: Ming Luo, Max S. Wicha, Jun-Lin Guan. Kinase-independent function of focal adhesion kinase in lung metastasis of breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1627.

Undiagnosed prostatic malignancy at the time of radical cystoprostatectomy after prior prostatic radiation therapy.

Purpose:We determined the prevalence of prostatic malignancy in patients undergoing radical cystoprostatectomy (RC) for urothelial carcinoma (UC) with a history of radiation therapy (XRT) treatment for prostatic adenocarcinoma (PCa).Materials and Methods:Fifty-three men who underwent a RC for UC that were previously treated for PCa with XRT were retrospectively identified. Pathology reports were reviewed to assess for residual PCa or prostatic UC at the time of surgery.Results:Thirteen (25%) patients had residual PCa, 16 (30%) had prostatic UC, and 8 (15%) had both. Sixteen (30%) patients had no evidence of prostatic disease. Patients with PCa had median tumor volume of 2.2 cc (interquartile range: 1.2–2.5 cc) and one-third had high-risk features (Gleason score >8 or pT3-T4 disease). Sixteen of 24 patients (67%) with prostatic UC had a stromal invasion, 5 (21%) had a ductal invasion, and 3 (13%) had carcinoma in situ. Tumors at bladder neck or trigone during transurethral resection were predictive of prostatic UC (odds ratio: 4.32, 95% confidence interval: 1.2–15.5, P = 0.025).Conclusions:Despite prior XRT for PCa, less than one-third of patients had no prostatic disease at the time of RC. Routine prostatic sampling should be considered in these patients especially if considering the orthotopic diversion.

Functional roles of MMP14 and MMP15 in early postnatal mammary gland development.

During late embryogenesis, mammary epithelial cells initiate migration programs that drive ductal invasion into the surrounding adipose-rich mesenchyme. Currently, branching morphogenesis is thought to depend on the mobilization of the membrane-anchored matrix metalloproteinases MMP14 (MT1-MMP) and MMP15 (MT2-MMP), which drive epithelial cell invasion by remodeling the extracellular matrix and triggering associated signaling cascades. However, the roles that these proteinases play during mammary gland development in vivo remain undefined. Here, we characterize the impact of global Mmp14 and Mmp15 targeting on early postnatal mammary gland development in mice. Unexpectedly, both Mmp14-/- and Mmp15-/- mammary glands retain the ability to generate intact ductal networks. Although neither proteinase is required for branching morphogenesis, transcriptome profiling reveals a key role for MMP14 and MMP15 in regulating mammary gland adipocyte differentiation. Whereas MMP14 promotes the generation of white fat depots crucial for energy storage, MMP15 differentially controls the formation of thermogenic brown fat. Taken together, these data not only indicate that current paradigms relevant to proteinase-dependent morphogenesis need be revisited, but also identify new roles for the enzymes in regulating adipocyte fate determination in the developing mammary gland.