bluuebunny/crossref_metadata_2025 huggingface dataset:
Ethyl Acetate Fraction from Leandra dasytricha (A. Gray) Cong. Leaves Promotes Vasodilatation and Reduces Blood Pressure in Normotensive and Hypertensive Rats
Authors: da Silva Rita de Cassia Vilhena , Bolda Mariano Luísa Nathália , Bidinha Eleine Renata , Bueno de Almeida Camila Leandra , Cechinel-Filho Valdir , Santos Zanuncio Vanessa Samudio , Silva Denise Brentan , Gasparotto Junior Arquimedes , de Souza Priscila
Published: November 2021

Leandra dasytricha (A. Gray) Cong. is widely distributed in the south of Brazil and is commonly used for cardiovascular and kidney ailments. For this study, we used male Wistar normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs) to verify the effects of the ethyl acetate fraction (EAF) obtained from L. dasytricha leaves on isolated aorta relaxation and in the arterial blood pressure. The EAF was analyzed by LC-DAD-MS, and several components were annotated, including hydrolysable tannins, triterpenes, and O- and C-glycosylated dihydrochalcones, such as the most intense ion peak relative to C-hexosyl phloretin (nothofagin; compound number 13). The EAF caused a concentration and endothelium-dependent relaxation of the aorta in both NTRs and SHRs. This effect was abolished in the endothelium-denuded aorta. L-NAME, a nonselective nitric oxide synthase inhibitor, and ODQ, a soluble guanylate cyclase inhibitor, entirely blocked the EAF-induced relaxation. The presence of a muscarinic receptor antagonist or a cyclooxygenase inhibitor did not alter the EAF's effectiveness in relaxing the aorta. The preincubation with tetraethylammonium, a Ca2+-activated K+ channel blocker, and with 4-aminopyridine, a voltage-dependent K+ channel blocker, significantly interfered with the EAF's relaxation. However, the incubation with glibenclamide, an ATP-sensitive K+ channel blocker, and barium chloride, an inward-rectifier K+ channel blocker, did not interfere with the EAF-induced relaxation. The EAF treatment also caused a dose-dependent decrease in the mean arterial pressure, systolic arterial pressure, and diastolic arterial pressure of both NTRs and SHRs, without significantly interfering with heart rate values. In conclusion, this study demonstrated the EAF-induced vasorelaxant and hypotensive actions, primarily dependent on the endothelium function and mainly with the participation of the nitric oxide and Ca2+-activated and voltage-dependent K+ channels.
Research on Failure Characteristics of Concrete Three-Point Bending Beams with Preexisting Cracks in Different Positions Based on Numerical Simulation
Authors: Zhao Liuqun , Zheng Li , Qin Hui , Geng Tiesuo , Tan Yonggang , Zhang Zhe
Published: November 2021

Concrete three-point bending beams with preexisting cracks are widely used to study the growth process of I-II mixed mode cracks. Studying the failure characteristics of preexisting cracks at different locations on concrete three-point bending beams not only has important scientific significance but also has a wide range of engineering application backgrounds in the safety assessment of engineering structures. In this paper, through several numerical experiments, the influence of preexisting cracks at different positions on the failure characteristics of concrete three-point bending beams is studied, and three typical failure modes are obtained. The failure process of the specimens with three typical failure modes is discussed in detail, and it is pointed out that the crack failure mode is tensile failure. The change trends of bearing capacity, acoustic emission quantity, and acoustic emission energy of three typical failure modes are analyzed. The maximum bearing capacity, the maximum acoustic emission quantity, and energy of three failure modes of concrete three-point bending beams generally show an increasing trend.
Aerobic Exercise Attenuates Pressure Overload-Induced Cardiac Dysfunction through Promoting Skeletal Muscle Microcirculation and Increasing Muscle Mass
Authors: Yuan Ling-Yan , Du Pei-Zhao , Wei Min-Min , Zhang Qi , Lu Le , Tian Xu , Fu Shao-ting , Zeng Xiao-Li
Published: November 2021

Background. Aerobic exercise has been proven to have a positive effect on cardiac function after hypertension; however, the mechanism is not entirely clarified. Skeletal muscle mass and microcirculation are closely associated with blood pressure and cardiac function. Objective. This study was designed to investigate the effects of aerobic exercise on the skeletal muscle capillary and muscle mass, to explore the possible mechanisms involved in exercise-induced mitigation of cardiac dysfunction in pressure overload mice. Methods. In this study, 60 BALB/C mice aged 8 weeks were randomly divided into 3 groups: control (CON), TAC, and TAC plus exercise (TAE) group and utilized transverse aortic constriction (TAC) to establish hypertensive model; meanwhile, treadmill training is used for aerobic exercise. After 5 days of recovery, mice in the TAE group were subjected to 10-week aerobic exercise. Carotid pressure and cardiac function were examined before mice were executed by Millar catheter and ultrasound, respectively. Muscle mass of gastrocnemius was weighed; cross-sectional area and the number of capillaries of gastrocnemius were detected by HE and immunohistochemistry, respectively. The mRNA and protein levels of VEGF in skeletal muscle were determined by RT-PCR and western blot, respectively. Results. We found that ① 10-week aerobic exercise counteracted hypertension and attenuated cardiac dysfunction in TAC-induced hypertensive mice; ② TAC decreased muscle mass of gastrocnemius and resulted in muscle atrophy, while 10-week aerobic exercise could reserve transverse aortic constriction-induced the decline of muscle mass and muscle atrophy; and ③ TAC reduced the number of capillaries and the protein level of VEGF in gastrocnemius, whereas 10-week aerobic exercise augmented the number of capillaries, the mRNA and protein levels of VEGF in mice were subjected to TAC surgery. Conclusions. This study indicates that 10-week aerobic exercise might fulfill its blood pressure-lowering effect via improving skeletal muscle microcirculation and increasing muscle mass.
PARP Inhibitors – Trapped in a Toxic Love Affair
Authors: Krastev Dragomir B. , Wicks Andrew J. , Lord Christopher J.
Published: November 2021

It is often the case that when an investigational cancer drug first enters clinical development, its precise mechanism of action is unclear. This was the case for PARP inhibitors (PARPi) used to treat homologous recombination–defective cancers. In 2012, nearly a decade after the first PARPi entered clinical development, work from Murai and colleagues demonstrated that clinical PARPi not only inhibit the catalytic activity of PARP1, PARylation, but also “trap” PARP1 on DNA; this latter effect being responsible for much of the tumor cell cytotoxicity caused by these drugs. We discuss how this work not only changed our understanding about how PARPi work, but also stimulated subsequent dissection of how PARP1 carries out its normal function in the absence of inhibitor.

See related article by Murai and colleagues, Cancer Res 2012;72:5588–5599.
Preclinical Characterization of a Next-Generation Brain Permeable, Paradox Breaker BRAF Inhibitor
Authors: Wichmann Jürgen , Rynn Caroline , Friess Thomas , Petrig-Schaffland Jeannine , Kornacker Martin , Handl Cornelia , Emmenegger Jasmin , Eckmann Jan , Herting Frank , Frances Nicolas , Hunziker Daniel , Krummenacher Daniela , Rüttinger Dominik , Ribeiro Alison , Bacac Marina , Brigo Alessandro , Hewings David S. , Dummer Reinhard , Levesque Mitchell P. , Schnetzler Gabriel , Martoglio Bruno , Bischoff James R. , Pettazzoni Piergiorgio
Published: November 2021

Purpose:

Disease progression in BRAF V600E/K positive melanomas to approved BRAF/MEK inhibitor therapies is associated with the development of resistance mediated by RAF dimer inducing mechanisms. Moreover, progressing disease after BRAFi/MEKi frequently involves brain metastasis. Here we present the development of a novel BRAF inhibitor (Compound Ia) designed to address the limitations of available BRAFi/MEKi.

Experimental Design:

The novel, brain penetrant, paradox breaker BRAFi is comprehensively characterized in vitro, ex vivo, and in several preclinical in vivo models of melanoma mimicking peripheral disease, brain metastatic disease, and acquired resistance to first-generation BRAFi.

Results:

Compound Ia manifested elevated potency and selectivity, which triggered cytotoxic activity restricted to BRAF-mutated models and did not induce RAF paradoxical activation. In comparison to approved BRAFi at clinical relevant doses, this novel agent showed a substantially improved activity in a number of diverse BRAF V600E models. In addition, as a single agent, it outperformed a currently approved BRAFi/MEKi combination in a model of acquired resistance to clinically available BRAFi. Compound Ia presents high central nervous system (CNS) penetration and triggered evident superiority over approved BRAFi in a macro-metastatic and in a disseminated micro-metastatic brain model. Potent inhibition of MAPK by Compound Ia was also demonstrated in patient-derived tumor samples.

Conclusions:

The novel BRAFi demonstrates preclinically the potential to outperform available targeted therapies for the treatment of BRAF-mutant tumors, thus supporting its clinical investigation.
Abstract PO-016: Directed evolution generates novel oncolytic H-1 parvoviruses with improved therapeutic efficacy in virus-resistant pancreatic cancer cells
Authors: Garcin Pierre , Kazemimanesh Monireh , Lulka Hubert , Dusetti Nelson , Labrousse Guillaume , Benuzzi Emilie , Buscail Louis , Cordelier Pierre
Published: November 2021

Despite considerable promise and emerging clinical success, several challenges impede the broader implementation of novel immunotherapies such as oncolytic virus(OV)-based gene therapy, including for patients with pancreatic cancer (PDAC). One of such challenge is inter-patient variability that may impact on OV selectivity and killing efficacy for tumor cells. For this study, we selected the rat parvovirus H-1 (H-1PV) that is nonpathogenic in humans and has a natural oncolytic activity in several cancer models. The safety and tolerability of H-1PV was recently demonstrated in early clinical trials for glioma and PDAC. However, we report here that H-1PV infection, oncolytic and pro-apoptotic activity are limited in PDAC cells, including patient-derived primary cells. To address this concern, we applied a directed evolution strategy to generate H-1PV variants with specific activity towards PDAC cells. Following selection using patient primary cells, we managed to isolate clonal, PDAC-adapted H-1PVs that induce PDAC cells lysis as compared to parental H-1PV while infection of normal pancreatic cells remained negligeable. Genome sequencing of the tumor adapted virus reveals mutations in promoting and viral capsid sequences. In vivo, the tumor-adapted H1PV demonstrates greater anti-tumor effect than parental H-1PV, following intravenous administration in an experimental model of orthotopic pancreatic tumors engrafted in immunodeficient mice. To our knowledge, we report here for the first time the production of highly selective and potent OV using directed evolution to override PDAC resistance to virotherapy. While the molecular mechanisms involved are still under investigation, this project is a first step towards precision medicine strategies based on OV.

Citation Format: Pierre Garcin, Monireh Kazemimanesh, Hubert Lulka, Nelson Dusetti, Guillaume Labrousse, Emilie Benuzzi, Louis Buscail, Pierre Cordelier. Directed evolution generates novel oncolytic H-1 parvoviruses with improved therapeutic efficacy in virus-resistant pancreatic cancer cells [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-016.
Abstract PO-037: Development of an RGD CRISPR-modified Clostridium novyi NT spores as an intravenous oncotherapy
Authors: Dailey Kaitlin M. , Vance Krysten , McAndrews Kyle , Jacobson Reed I. , Delgado Jandro , Johnson Paige R. , Woolery Taylor M. , Orr Megan , Kim Jiha , Mallik Sanku , Bayles Kenneth W. , Hollingsworth Michael A. , Brooks Amanda E.
Published: November 2021

The efficacy of current oncotherapeutics is largely limited by an inability to access avascular tissues, which is in part responsible for forty years of stagnant pancreatic cancer statistics where the median survival remains a mere six months. Oncolytic bacteria such as Clostridium novyi-NT overcome this challenge with its ultrasensitive, innate affinity for hypoxic/necrotic areas found at the center of solid tumors and their metastases. While preclinical and clinical data from intratumoral injections of C. novyi-NT are promising, many tumors are inaccessible to such injections. Preclinical trials of analogous IV injections have uncovered other obstacles such as rapid clearance of C. novyi-NT by the immune system independent of septic complications. To mitigate rapid clearance, CRISPR/Cas9n was used to genetically modify a non-toxic form of C. novyi-NT to express a tumor targeting RGD peptide on the spore surface. Through this novel, first of its kind, methodology, spores with stronger affinity to a surface coated with the targeted binding partner of RGD, aVb3 integrin, have been generated. Importantly, there was no statistically significant difference in the genetically modified spore’s capacity for sporulation or germination when compared to unmodified C. novyi-NT spores, nor was a difference in lytic capacity observed, suggesting no relevant off-target effects from genomic modification. Biodistribution and efficacy of non-toxic RGD-modified spores was evaluated in an immunocompetent, syngeneic, pancreatic cancer murine model. Ongoing efforts to characterize the biodistribution and efficacy of the intravenously injected RGD-modified C. novyi-NT include the application of multiplex immunofluorescence, laser microdissection, and live, whole animal imaging. Supported as a pilot project by funds from NIH COBRE grant 1P20GM109024, Doctoral Dissertation Funds to KMD from NDSU, and by discretionary funds from investigators at UNMC.

Citation Format: Kaitlin M. Dailey, Krysten Vance, Kyle McAndrews, Reed I. Jacobson, Jandro Delgado, Paige R. Johnson, Taylor M. Woolery, Megan Orr, Jiha Kim, Sanku Mallik, Kenneth W. Bayles, Michael A. Hollingsworth, Amanda E. Brooks. Development of an RGD CRISPR-modified Clostridium novyi NT spores as an intravenous oncotherapy [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-037.
Abstract PO-038: LAMC2: New player in stemness and tumor progression in pancreatic cancer
Authors: Cave Donatella Delle , Iavazzo Tea Teresa , Mangini Maria , Andolfi Gennaro , Pirozzi Teresa , Di Domenico Annalisa , De Luca Annachiara , Lonardo Enza
Published: November 2021

Pancreatic ductal adenocarcinoma (PDAC) is a devastating and essentially incurable disease, typically characterized by high chemoresistance and metastatic spread attributable to cancer stem cells (CSCs). This subpopulation is critical for tumor initiation and recurrence, but the mechanism through which they acquire metastatic traits is not well understood. Hence, targeting the CSC niche and their plasticity could be a complementary therapeutic strategy against cancer. Laminin subunit-γ-2 (LAMC2) is an epithelial basement membrane protein, which controls cell motility and adhesion and is widely expressed in the majority of human tumors. However, its role in PDAC remains largely unknown. In several patient cohorts we observed that high levels of LAMC2 significantly correlated with shorter overall survival. In addition, the tissue microarray analysis on PDAC sections revealed prognostic significance of LAMC2 expression in tumor with high grade of aggressiveness (i.e., G2 and G3). To determine the role of LAMC2 in sustaining tumorigenicity, we knocked down it in patient-derived xenografts (PDX) cells using lentiviral shRNA constructs. The silencing of LAMC2 resulted in decreased self-renewal, invasiveness, tumorigenicity and gemcitabine resistance both in vitro and in vivo. To identify or track the LAMC2 tumor cell population in an intact environment we engineered primary PDAC cells that carry EGFP cassette knocked in the LAMC2 locus through the CRISPR-Cas9 technique. Analysis of LAMC2-EGFP+ cells isolated from tumor demonstrated that these cells express a gene program similar to that of highly metastatic stem cells and that they initiate and propagate both the primary tumor and the metastasis to recipient mice very efficiently compared to their counterpart. In conclusion, we identified a highly metastatic subpopulation of cancer stem cells, characterized by high levels of LAMC2. Strategies aimed at targeting the LAMC2 population may be effective in reducing tumor aggressiveness in combination with conventional therapy.

Citation Format: Donatella Delle Cave, Tea Teresa Iavazzo, Maria Mangini, Gennaro Andolfi, Teresa Pirozzi, Annalisa Di Domenico, Annachiara De Luca, Enza Lonardo. LAMC2: New player in stemness and tumor progression in pancreatic cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-038.
Abstract PO-051: PANOVA-3: A phase III study of tumor treating fields with nab-paclitaxel and gemcitabine for front-line treatment of locally advanced pancreatic adenocarcinoma
Authors: Picozzi Vincent J. , Macarulla Teresa , Philip Philip A. , Becerra Carlos R. , Dragovich Tomislav
Published: November 2021

Tumor Treating Fields (TTFields) are a non-invasive, loco-regional antimitotic treatment modality, approved for the treatment of glioblastoma and malignant pleural mesothelioma. TTFields at a specific frequency (150-200 kHz) are delivered via arrays placed on the skin surrounding the tumor site. TTFields predominantly act by disrupting the formation of the mitotic spindle during metaphase. TTFields were effective in multiple preclinical models of pancreatic cancer. The Phase 2 PANOVA study, the first trial testing TTFields in pancreatic cancer patients, demonstrated the safety and preliminary efficacy of TTFields when combined with nab-paclitaxel and gemcitabine in both metastatic and locally advanced pancreatic adenocarcinoma (LAPC). The Phase 3 PANOVA-3 trial (NCT03377491) is designed to test the efficacy and safety of adding TTFields to nab-paclitaxel and gemcitabine combination in LAPC. Patients (N = 556) with unresectable, LAPC (per NCCN guidelines) will be enrolled in this prospective, randomized trial. Patients should have an ECOG score of 0-2 and no prior progression or treatment. Patients will be stratified based on their performance status and geographical region, and will be randomized 1:1 to TTFields plus nab-paclitaxel and gemcitabine or to nab-paclitaxel and gemcitabine alone. Chemotherapy will be administered at standard dose of nab-paclitaxel (125 mg/m2) and gemcitabine (1000 mg/m2) on day 1, 8, and 15 of a 28 day cycle. The protocol has recently been amended to incorporate the use of a smaller, lighter (weight reduced from 6 to 2.7 lbs.) TTFields device. TTFields (150 kHz) will be delivered at least 18 hours/day until local disease progression per RECIST Criteria V1.1. Follow up will be performed q4w, including a CT scan of the chest and abdomen q8w. Following local disease progression, patients will be followed monthly for survival. Overall survival will be the primary endpoint. Progression-free survival, objective response rate, rate of resectability, quality of life, and toxicity will all be secondary endpoints. Sample size was calculated using a log-rank test comparing time to event in patients treated with TTFields plus chemotherapy with control patients on chemotherapy alone. PANOVA-3 is designed to detect a hazard ratio 0.75 in overall survival. Type I error is set to 0.05 (two-sided) and power to 80%.

Citation Format: Vincent J. Picozzi, Teresa Macarulla, Philip A. Philip, Carlos R. Becerra, Tomislav Dragovich. PANOVA-3: A phase III study of tumor treating fields with nab-paclitaxel and gemcitabine for front-line treatment of locally advanced pancreatic adenocarcinoma [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-051.
Abstract PO-057: Targeting ErbB2 degradation via the ubiquitin–proteasome pathway to inhibit the metastasis of pancreatic cancer
Authors: Zhang Bo , Teng Fei , Lin Nengming
Published: November 2021

Objective: Pancreatic cancer is currently one of the most lethal human cancers and continues to be a major unsolved health problem in the 21st century. Since pancreatic cancer has a high metastatic potential, chemotherapy is inevitable for most pancreatic cancer patients, whereas the clinical responses to anticancer agents are not satisfied. In this work, we designed and synthesized a novel compound, termed as 6-MPA, and found that 6-MPA could improve the proteasomal degradation of ErbB2 pancreatic cancer cells. Methods: The inhibitory effects of 6-MPA on migration and invasion was measured by scratch experiment, wound healing assay and transwell experiment. Clone formation and CCK-8 assay was used to determine the cytotoxicity of 6-MPA. Real-time PCR and western blotting was used to determine the expression of indicated genes or proteins respectively. Immunoprecipitation was used to characterize protein–protein interactions. Results: 4, 8, and 16 μM of 6-MPA exhibited an inhibitory effects on the migration of SW1990 cells and the inhibition rates reached 40.2±10.1%, 58.2±11.2%, and 65.2±9.9% in vitro, respectively. RNA-seq showed that 6-MPA treatment manipulated ErbB2 level and its downstream PI3K/AKT/mTOR signaling pathways, which was verified by western blotting assay. Treatment with 6-MPA could enhance the interaction between ErbB2 and ubiquitin, and thus promote the proteasomal degradation of ErbB2. Conclusion: In this study, we obtained a novel small molecule 6-MPA, which could specifically degrade ErbB2 through the activation of protein ubiquitinination. And we found that 6-MPA possessed potent inhibitory effects on migration and invasion of pancreatic cancer cells. Therefore, our work illustrated a novel structure skeleton that could be used to target ErbB2, and potentially suppressed the metastasis of pancreatic cancer.

Citation Format: Bo Zhang, Fei Teng, Nengming Lin. Targeting ErbB2 degradation via the ubiquitin–proteasome pathway to inhibit the metastasis of pancreatic cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-057.
Abstract PO-076: Murine adapted FOLFIRINOX for standard-of-care in KPC mice
Authors: Whittle Martin C. , Palkar Aditi , Fasnacht Rachael , Yan James , Kheng Borith , Sinfuego Kianna , Thorsen Shelley , Hingorani Sunil R.
Published: November 2021

Until relatively recently, the standard-of-care (SOC) treatment for pancreas cancer was not appreciably superior to best supportive care in prolonging life, and potentially new treatment strategies had a relatively modest bar to surmount. New combination regimens have emerged and novel therapies should therefore be compared against the current SOC. However, in preclinical development of novel agents, efficacy is frequently compared against vehicle control and less commonly against cytotoxic chemotherapy. Furthermore, virtually all studies of cytotoxic chemotherapy for PDA in mice have used gemcitabine (gem) monotherapy as standard-of-care, although this is no longer the standard for this disease. One of the current standard regimens, gem+nab-paclitaxel, cannot be given in mice without inducing immune responses because of the incorporation of human albumin in developing the taxol nanoparticles. The other new standard, FOLFIRINOX, is a complex 4-agent regimen that includes a 46-hour continuous infusion component that has been prohibitively challenging to deliver to animals. We have now built an appropriate dosing regimen incorporating these 4 agents that required a deliberate, step-by-step, program of empiric testing of the drugs, first, one at a time, then in various doublets and triplets, and, finally, as a whole regimen. The studies included close clinical evaluations and serial blood tests to follow complete blood counts (CBC). We have incorporated this regimen as the new SOC in our Murine Clinical Trials Program (MCTP). We propose that this mur-FOLIFIRINOX regimen should represent the new standard for murine studies.

Citation Format: Martin C. Whittle, Aditi Palkar, Rachael Fasnacht, James Yan, Borith Kheng, Kianna Sinfuego, Shelley Thorsen, Sunil R. Hingorani. Murine adapted FOLFIRINOX for standard-of-care in KPC mice [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-076.
Abstract PO-092: Influence of the IL-13-receptor alpha 1 chain on the malignant phenotype of pancreatic cancer cells
Authors: Shi Jingwei , Kornmann Marko , Traub Benno
Published: November 2021

Introduction: Increasing evidence indicates salient activities of Interleukin (IL)-4, IL-13 and their specific Type-II-IL-4 receptor complex (IL-4Rα/IL-13Rα1) in carcinomas including pancreatic cancer. While IL-4 seems to be tumor-promoting in PDAC, the specific role of the IL-13/IL-13Rα1 axis was yet to be determined. Both cytokines are abundant in PDAC through cells of the tumor microenvironment. In this project, we planned to determine the specific effects of the IL-13Rα1-receptor chain on the malignant phenotype of pancreatic cancer cells. Methods: Expression of IL-13Rα1, IL-4Rα, and the human common gamma chain (γc) in cultured pancreatic cancer cell lines AsPC-1, Capan-1, PANC-1, MIA PaCa-2 and A818-6 as well as downstream signal transduction factors of IL13-Rα1 were detected by Western blot. The role of IL-13Rα1 in pancreatic cancer cell proliferation, mobility and migration was demonstrated based on the successful establishment of stable shRNA based IL-13Rα1-knockdown (KD) clones in Capan-1 and MIA PaCa-2 cell lines. Results: Not only the basal anchorage-dependent growth but also the anchorage-independent growth in soft agar were inhibited in Capan-1 and MIA PaCa-2 cells due to IL-13Rα1-downregulation. Mechanistically, IL-13Rα1-KD was associated with increased apoptosis. Additionally, signal transduction factors STAT3, STAT6, PI3K, Akt and ERK1/2 were activated in Capan-1 exposed to exogenous IL-4 and IL-13. Interestingly, IL-13Rα1-KD reduced the basal expression of STAT3, Akt and ERK1/2. IL-4 and IL-13 induced activation of STAT3, ERK1/2, Akt and STAT6 was significantly suppressed in Capan-1-IL-13Rα1-KD (C-KD) clones. Furthermore, IL-13Rα1-KD resulted in enhanced mobility and migration in Capan-1 cells. Conclusion: It was demonstrated that endogenous IL-13Rα1 is vital for pancreatic cancer cell growth with the involvement of STAT3/6, ERK1/2 and Akt. Surprisingly, IL-13Rα1-KD resulted in enhanced mobility and migration. Overall, IL-13Rα1 plays a critical but heterogenous role in PDAC. Our findings increase the understanding of the different functions and mechanisms involving IL-13Rα1 in pancreatic cancer progression.

Citation Format: Jingwei Shi, Marko Kornmann, Benno Traub. Influence of the IL-13-receptor alpha 1 chain on the malignant phenotype of pancreatic cancer cells [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-092.
Abstract PO-101: Characterization of longitudinally collected fine needle aspiration biopsies of pancreatic ductal adenocarcinoma upon endoscopic ultrasound guided radiofrequency ablation
Authors: Desai Krisha , Lawrence Patrick Varun , Wadsworth Christopher , Mangal Nagina , Habib Nagy , Sadanandam Anguraj , Sodergren Mikael
Published: November 2021

Background: Most patients with pancreatic ductal adenocarcinoma (PDAC) are metastatic at presentation with dismal prognosis warranting improved systemic therapy options. Longitudinal sampling for assessment of treatment response poses a challenge for validating novel therapies. In this proof-of-principle study, we evaluate the role of endoscopic ultrasound (EUS)-guided serial fine-needle aspiration biopsies (FNABs) to study the mechanism of action of radiofrequency ablation (RFA). Methods: Patients with stage III inoperable PDAC with prior exposure to gemcitabine were selected into ARDEO (ethically approved Phase-II prospective clinical study of EUS-RFA). Post examination, targeted RF was delivered thrice and sequential FNABs of tumor were taken before and after treatment. Transcriptomic profiling of 6 FNABs from 2 patients was performed using a custom NanoString gene panel (144 genes) consisting of cancer and cancer-associated fibroblast (CAFs) subtypes along with immune changes. CAF culture was established from one FNAB and characterised by immunofluorescence and immunoblotting. Results: RFA treatments were well tolerated without any complications and both patients had stable disease immediately after EUS-RFA. Two-course RFA led to upregulation of CD1E gene (participates in antigen presentation) in both, patient 1 and 2 (4.5 and 3.9-fold) compared to baseline. Patient 1 showed increased expression of T cell genes (CD4 – 8.7-fold, CD8 – 35.7-fold), cytolytic function (6.4-fold) and inflammatory response (8-fold) post-RFA. Greater than 2-fold upregulation of CD274 (PDL1), IDO1, PDCD1 and TNFRSF18 (GITR) was observed post 2nd RFA in both the patients. Further, two-course RFA led to increased PDGFRa (4.5 and 9-fold) in both patients along with enrichment of pCAF subtypes B and C in patient 1 and subtypes A, B and D in patient 2. Immunofluorescence staining revealed expression of PDGFRa, aSMA, VIM, POSTN and MYH11 on patient2-derived CAFs post 1st RFA; validated by immunoblotting. Finally, RFA led to downregulation of classical PDA subtype in both patients. Conclusions: This feasibility study validates longitudinal sampling by EUS-FNABs as an appropriate research tool to study tumor microenvironmental changes associated with local pancreatic immunomodulatory techniques like RFA.

Citation Format: Krisha Desai, Patrick Varun Lawrence, Christopher Wadsworth, Nagina Mangal, Nagy Habib, Anguraj Sadanandam, Mikael Sodergren. Characterization of longitudinally collected fine needle aspiration biopsies of pancreatic ductal adenocarcinoma upon endoscopic ultrasound guided radiofrequency ablation [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-101.
Abstract PO-116: Deletion of Arginase 1 in myeloid cells alters the pancreatic cancer microenvironment
Authors: Menjivar Rosa E. , Nwosu Zeribe , Du Wenting , Donahue Katelyn , Espinoza Carlos , Velez-Delgado Ashley , Brown Kristee , Yan Wei , Halbrook Christopher , Zhang Yaqing , Lyssiotis Costas , Pasca di Magliano Marina
Published: November 2021

Pancreatic ductal adenocarcinoma (PDA) is a deadly disease with a 5-year survival of only 10%. PDA is characterized by an abundant fibroinflammatory stroma that includes abundant fibroblasts and immune cells, mainly myeloid cells. Infiltrating myeloid cells express high levels of Arginase 1 (Arg1), an enzyme that metabolizes L-arginine. Conversely, CD8+ T cells are scarce in PDA, and when present have an overwhelmingly exhausted phenotype. Whether myeloid cell Arginase is a key driver of immune suppression in pancreatic cancer is unknown. Here, we tested the hypothesis that myeloid cells in the tumor microenvironment mediate immune suppression in PDA through expression of Arg1. To test this hypothesis, we used a combination of genetically engineered pancreatic cancer mouse models and pharmacological approaches. Using a FlpO- and Cre-based dual recombinase system, we have generated a mouse model that develops pancreatic cancer spontaneously because of oncogenic Kras expression in the epithelium, while at the same time lacking Arg1 expression in the myeloid cell compartment (Ptf1a-FlpO/+;KrasFrt-STOP-Frt-G12D/+;LysMCre;Arg1f/f). To complement the genetic model and inhibit the function of Arginase systemically, we used an Arginase inhibitor (CB-1158, Incyte, Inc.) alone and in combination with an immune checkpoint blockade (anti-PD1). Using these multiple approaches, we observed decrease progression to invasive disease in the genetic model, and sensitization to immune checkpoint treatment in the transplantation model. In both settings, changes in tumor growth were accompanied by an increase in CD8+ T cell infiltration and activation. These changes support the notion that myeloid Arg1 is mediator of immune suppression in PDA, and a potential therapeutic target.

Citation Format: Rosa E. Menjivar, Zeribe Nwosu, Wenting Du, Katelyn Donahue, Carlos Espinoza, Ashley Velez-Delgado, Kristee Brown, Wei Yan, Christopher Halbrook, Yaqing Zhang, Costas Lyssiotis, Marina Pasca di Magliano. Deletion of Arginase 1 in myeloid cells alters the pancreatic cancer microenvironment [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-116.
Abstract PO-127: A uPA/uPAR axis in both the tumor cell and stromal compartment drives PDAC disease progression
Authors: Yang Yi , Abrahams Sara R. , Kothari Aditi , Matada Harshi , Davey Keely , Wolberg Alisa S. , Flick Matthew J.
Published: November 2021

Pancreatic ductal adenocarcinoma (PDAC) is a lethal solid tumor malignancy with a 5-year survival rate of 9%. In both patients and animal models of disease, PDAC is associated with robust coagulation system activity. Intriguingly, in addition to being a rich source of procoagulant factors, PDAC tumors highly express fibrinolytic system components. Supporting this concept, urokinase plasminogen activator (uPA) and uPA receptor (uPAR) expression positively correlates with reduced overall patient survival. Here, we tested the hypothesis that the expression and activity of plasminogen activation (PA) system components are functionally linked to PDAC tumor growth and disease progression. We generated C57Bl/6-derived KPC (i.e., KRasG12D, TRP53R172H) PDAC cell lines in which uPA and uPAR were knocked out using CRISPR-Cas9. We then analyzed orthotopic tumor growth and experimental metastasis in mice carrying null or functional mutations in uPA, uPAR, or plasminogen to evaluate the interplay of PA components derived from tumor cells and/or stromal cells in mediating PDAC progression. Although both KPC cell CRISPR variants retained procoagulant function, elimination of tumor cell uPA or uPAR yielded significantly smaller tumors when compared to Cas9 control tumor cells in wildtype mice. Similarly, the growth of WT KPC tumor cells in C57Bl/6 background uPA-KO or uPAR-KO mice also resulted in reduced tumor growth. To our surprise, the metastasis potential of WT KPC tumor cells in uPA-KO or uPAR-KO mice did not change when compared to wildtype mice. Regarding to the uPA/uPAR axis downstream effector plasminogen, the growth of WT KPC tumors in plasminogen-KO mice was also significantly reduced, but not to the same extent as when eliminating uPA or uPAR. In addition, eliminating plasminogen drastically reduced WT KPC tumor cells metastasis potential. In conclusion, our data suggest a mechanism whereby uPA functions through uPAR in both the tumor cell and stromal cell compartments to promote PDAC progression through plasminogen-dependent and -independent mechanisms.

Citation Format: Yi Yang, Sara R. Abrahams, Aditi Kothari, Harshi Matada, Keely Davey, Alisa S. Wolberg, Matthew J. Flick. A uPA/uPAR axis in both the tumor cell and stromal compartment drives PDAC disease progression [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-127.
Association of Race and Ethnicity and Anticoagulation in Patients With Atrial Fibrillation Dually Enrolled in Veterans Health Administration and Medicare: Effects of Medicare Part D on Prescribing Disparities
Authors: Essien Utibe R. , Kim Nadejda , Magnani Jared W. , Good Chester B. , Litam Terrence M.A. , Hausmann Leslie R.M. , Mor Maria K. , Gellad Walid F. , Fine Michael J.
Published: November 2021

Background:

Racial and ethnic disparities in anticoagulation exist in atrial fibrillation management in Medicare and the Veterans Health Administration, but the influence of dual Veterans Health Administration and Medicare enrollment is unclear. We compared anticoagulant initiation by race and ethnicity in dually enrolled patients and assessed the role of Medicare part D enrollment on anticoagulation disparities.

Methods:

We identified patients with incident atrial fibrillation (2014–2018) dually enrolled in Veterans Health Administration and Medicare. We assessed any anticoagulant initiation (warfarin or direct-acting oral anticoagulants [DOACs]) within 90 days of atrial fibrillation diagnosis and DOAC use among anticoagulant initiators. We modeled anticoagulant initiation, adjusting for patient, provider, and facility factors, including main effects for race and ethnicity and Medicare part D enrollment and an interaction term for these variables.

Results:

In 43 789 patients, 8.9% were Black, 3.6% Hispanic, and 87.5% White; 10.9% participated in Medicare part D. Overall, 29 680 (67.8%) patients initiated any anticoagulant, of whom 17 568 (59.2%) initiated DOACs. Lower proportions of Black (65.2%) than Hispanic (67.6%) or White (68.0%) patients initiated any anticoagulant (
P
=0.001) and, lower proportions of Black (56.3%) and Hispanic (55.9%) than White (59.6%) patients (
P
=0.001) initiated DOACs. Compared with White patients, Black patients had significantly lower initiation of any anticoagulant (adjusted odds ratio, 0.89 [95% CI, 0.82–0.97]). The adjusted odds ratios for DOAC initiation were significantly lower for Black (0.72 [95% CI, 0.65–0.81]) and Hispanic (0.84 [95% CI, 0.70–1.00]) than White patients. The interaction between race and ethnicity and Medicare part D enrollment was nonsignificant for any anticoagulant (
P
=0.99) and DOAC (
P
=0.27) therapies.

Conclusions:

In dually enrolled Veterans Health Administration and Medicare patients with atrial fibrillation, Black patients were less likely to initiate any anticoagulant, and Black and Hispanic patients were less likely to initiate DOACs. Medicare part D enrollment did not moderate the associations between race and ethnicity and anticoagulant therapies.
Intravenous Thrombolysis Before Mechanical Thrombectomy for Acute Ischemic Stroke: A Meta‐Analysis
Authors: Du Houwei , Lei Hanhan , Ambler Gareth , Fang Shuangfang , He Raoli , Yuan Qilin , Werring David J. , Liu Nan
Published: November 2021

Background

Whether intravenous thrombolysis before mechanical thrombectomy provides additional benefit for functional outcome in acute ischemic stroke remains uncertain. We performed a meta‐analysis to compare the outcomes of direct mechanical thrombectomy (dMT) to mechanical thrombectomy with bridging using intravenous thrombolysis (bridging therapy [BT]) in patients with acute ischemic stroke.

Methods and Results

We performed a literature search in the PubMed, Excerpta Medica database, and Cochrane Central Register of Controlled Trials from January 1, 2003, to April 26, 2021. We included randomized clinical trials and observational studies that reported the 90‐day functional outcome in patients with acute ischemic stroke undergoing dMT compared with BT. The 12 included studies (3 randomized controlled trials and 9 observational studies) yielded 3924 participants (mean age, 68.0 years [SD, 13.1 years]; women, 44.2%; 1887 participants who received dMT and 2037 participants who received BT). A meta‐analysis of randomized controlled trial and observational data revealed similar 90‐day functional independence (odds ratio [OR], 1.04; 95% CI, 0.90–1.19), mortality (OR, 1.03; 95% CI, 0.78–1.36), and successful recanalization (OR, 0.93; 95% CI, 0.76–1.14) for patients treated with dMT or BT. Compared with those in the BT group, patients in the dMT group were less likely to experience symptomatic intracranial hemorrhage (OR, 0.68; 95% CI, 0.51–0.91;
P
=0.008) or any intracranial hemorrhage (OR, 0.71; 95% CI, 0.61–0.84;
P
<0.001).

Conclusions

In this meta‐analysis of patients with acute ischemic stroke, we found no significant differences in 90‐day functional outcome or mortality between dMT and BT, but a lower rate of symptomatic intracranial hemorrhage for dMT. These findings support the use of dMT without intravenous thrombolysis bridging therapy.

Registration

URL:
https://www.crd.york.ac.uk/prospero/
; Unique identifier: 42021234664.
Morphology and molecular phylogeny reveal two new species in Russula sect. Ingratae from China
Authors: CHEN BIN , SONG JIE , ZHANG JIN-HUA , LIANG JUN-FENG
Published: November 2021

Two new species of Russula are described and illustrated in this paper. Russula clavulus is recognised by a pale yellow pileus centre, white margin with tuberculate striation, white to pale lamellae with small pale yellow spots, white to light yellow spore print, subglobose to broadly ellipsoid spores with short or long ridges and hymenial cystidia on lamellae sides that are mainly subclavate or fusiform. Russula multilamellula is morphologically characterised by the brownish orange to hazel pileus centre and satin white to yellowish-white margin with brownish tinge, lamellulae that are usually irregular in length and often anastomosing with lamellae, subglobose to broadly ellipsoid spores with short or long ridges and clavate hymenial cystidia. The combination of morphological features and multigene phylogenetic analyses of ITS-nrLSU-RPB2-mtSSU data indicated that these two new taxa belong to Russula subg. Heterophyllidia sect. Ingratae.
Reorganizing parasitic Delesseriaceae: taxonomic revision of Asterocolax
Authors: FREESE JILLIAN M. , LANE CHRISTOPHER E.
Published: November 2021

Red algal parasites are highly host specific organisms that are morphologically reduced with decreased pigmentation. Only found within the Florideophyceae, red algal parasites have evolved within eight orders, with the greatest parasite diversity found in the Ceramiales. A quarter of the ceramialian parasites in the family Delesseriaceae are described in the genus Asterocolax. The initial morphological description of Asterocolax led to the creation of an independent genus devoted to these parasitic species, but molecular data have repeatedly demonstrated that Asterocolax species, and likely many other red algal parasites, resolve within the genera of free-living red algae, often within the same genus as their hosts. Here and in previous studies, phylogenetic analysis of the internal transcribed spacer region of the ribosomal DNA has shown at least six instances of independent evolution of Asterocolax species, mainly interspersed among the free-living macroalgal genera of Phycodrys and Polyneura. As most Asterocolax are sister species to their hosts, they are independently derived from a photosynthetic ancestor, and together, do not form a monophyletic parasitic genus. Here we conduct a long overdue taxonomic revision of the red algal parasitic genus Asterocolax, describe a new species within the genus, and propose nomenclatural changes for four other species to reduce the polyphyletic clades encompassing Asterocolax.
Complex Effects of Telecouplings on Forest Dynamics: An Agent-Based Modeling Approach
Authors: Yang Hongbo , Ligmann-Zielinska Arika , Dou Yue , Chung Min Gon , Zhang Jindong , Liu Jianguo
Published: November 2021

Rural areas are increasingly subject to the effects of telecouplings (socioeconomic and environmental interactions over distances) whereby their human and natural dynamics are linked to socioeconomic and environmental drivers operating far away, such as the growing demand for labor and ecosystem services in cities. Although there have been many studies evaluating the effects of telecouplings, telecouplings in those studies were often investigated separately, and how telecouplings may interact and affect dynamics of rural coupled human and natural systems (CHANS) jointly was rarely evaluated. In this study, we developed an agent-based model and simulated the impacts of two globally common telecouplings, nature-based tourism and labor migration, on forest dynamics of a rural CHANS, China’s Wolong Nature Reserve (Wolong). Nature-based tourism and labor migration can facilitate forest recovery, and the predicted forest areas in Wolong in 2030 would be reduced by 26.2 km2 (6.8%) and 23.9 km2 (6.2%), respectively, without their effects. However, tourism development can significantly reduce the probability of local households to have member(s) outmigrate to work in cities and decrease the positive impact of labor migration on forest recovery. Our simulations show that the interaction between tourism and labor migration can reduce the potential forest recovery by 3.5 km2 (5.0%) in 2030. Our study highlights that interactions among different telecouplings can generate significant impacts on socioeconomic and environmental outcomes and should be jointly considered in the design, management, and evaluation of telecouplings for achieving sustainable development goals.

Significance Statement

Rural areas are increasingly connected with other places through telecouplings, such as tourism and labor migration. However, telecouplings’ effects were often evaluated separately, and their interaction remains poorly understood. In this study, we evaluated how two globally common telecouplings, tourism and labor migration, jointly affect forest dynamics in a demonstration site using an agent-based modeling approach. Although both tourism and labor migration can benefit forest conservation, we found that their interaction generates an antagonistic effect: households’ involvement in tourism activities reduces their probability to have members outmigrate to work in cities and significantly diminishes the beneficial impact of labor migration on forest recovery. Our study highlights the importance of considering interaction among telecouplings in the management of telecouplings for sustainability.