EMT and Treatment Resistance in Pancreatic Cancer

Pancreatic cancer (PC) is the third leading cause of adult cancer mortality in the United States. The poor prognosis for patients with PC is mainly due to its aggressive course, the limited efficacy of active systemic treatments, and a metastatic behavior, demonstrated throughout the evolution of the disease. On average, 80% of patients with PC are diagnosed with metastatic disease, and the half of those who undergo surgery and adjuvant therapy develop liver metastasis within two years. Metastatic dissemination is an early event in PC and is mainly attributed to an evolutionary biological process called epithelial-to-mesenchymal transition (EMT). This innate mechanism could have a dual role during embryonic growth and organ differentiation, and in cancer progression, cancer stem cell intravasation, and metastasis settlement. Many of the molecular pathways decisive in EMT progression have been already unraveled, but little is known about the causes behind the induction of this mechanism. EMT is one of the most distinctive and critical features of PC, occurring even in the very first stages of tumor development. This is known as pancreatic intraepithelial neoplasia (PanIN) and leads to early dissemination, drug resistance, and unfavorable prognosis and survival. The intention of this review is to shed new light on the critical role assumed by EMT during PC progression, with a particular focus on its role in PC resistance

Understanding Patient Experience in Biliary Tract Cancer: A Qualitative Patient Interview Study

Cáncer del tracto biliar; Estudio de entrevista; Investigación cualitativaCàncer del tracte biliar; Estudi d'entrevista; Recerca qualitativaBiliary tract cancer; Interview study; Qualitative researchIntroduction Patients living with biliary tract cancer (BTC) experience a decline in health-related quality of life (HRQoL). This study aimed to obtain a comprehensive understanding of the patient experience of BTC-related signs/symptoms and the impacts of these on daily functioning and HRQoL. Methods Patients with BTC participated in qualitative semi-structured concept elicitation interviews. Signs/symptoms and impacts of BTC were initially explored by targeted literature searches and interviews with five clinicians. Patient interviews were transcribed and coded using qualitative research software. Concept saturation was assessed over five interview waves. A sign/symptom or impact was defined as “salient” if mentioned by ≥ 50% of patients, with a mean disturbance rating of ≥ 5 (0–10 scale). A conceptual model of the patient experience of BTC-related signs/symptoms and impacts was produced. Results Twenty-three patients from the USA (78% women; median age: 54 years), diagnosed as having early (n = 3), locally advanced (n = 11) or metastatic (n = 9) disease, were interviewed. Sixty-six signs/symptoms and 12 impacts were identified. Of these, 46 signs/symptoms and 8 impacts were not identified from the targeted literature or clinician interviews. Concept saturation was reached by the fourth of five interview waves. Fourteen disease-related signs/symptoms (including fatigue/lack of energy, abdominal pain, lack of appetite, insomnia and diarrhoea) and three impacts (physical, emotional and cognitive impacts) were deemed “salient”. The conceptual model included 50 signs/symptoms and 12 impacts. Conclusion Patients with BTC reported a range of signs/symptoms and impacts that negatively affect daily functioning and HRQoL.This project was funded by AstraZeneca, Gaithersburg, MD, USA. Research and publication fees are funded by the study sponsor

Diagnosis and treatment of cholangiocarcinoma in Italy: A Delphi consensus statement

Background: Clinical practice guidelines for the management of cholangiocarcinoma (CCA)/biliary tract cancer recommend genomic profiling to guide treatment decisions. Variable access to such profiling across Italy means many oncologists are unfamiliar with when and how to conduct genetic testing and prescribe targeted treatments. Methods: A Scientific Board of Italian oncologists who treat CCA (the authors) developed recommendations, based on recent clinical evidence, for using molecular testing in diagnosing, assessing, and treating CCA in Italy. The Delphi process was used to reach consensus on these recommendations among 38 Italian oncologists. Consensus was considered to be met if >= 66.7 % of the panel agreed or strongly agreed with each statement. Findings: Consensus was reached on 28 statements across four themes: (1) epidemiology and risk factors; (2) diagnosis, including molecular diagnosis; (3) treatment selection; and (4) treatment safety. Interpretation: These recommendations should aid Italian clinicians in selecting appropriate treatment options for their patients

NALIRIFOX versus nab-paclitaxel and gemcitabine in treatment-naive patients with metastatic pancreatic ductal adenocarcinoma (NAPOLI 3): a randomised, open-label, phase 3 trial

Gemcitabine; Metastatic pancreatic ductal adenocarcinomaGemcitabina; Adenocarcinoma ductal de pàncrees metastàticGemcitabina; Adenocarcinoma ductal de páncreas metastásicoBackground Pancreatic ductal adenocarcinoma remains one of the most lethal malignancies, with few treatment options. NAPOLI 3 aimed to compare the efficacy and safety of NALIRIFOX versus nab-paclitaxel and gemcitabine as first-line therapy for metastatic pancreatic ductal adenocarcinoma (mPDAC). Methods NAPOLI 3 was a randomised, open-label, phase 3 study conducted at 187 community and academic sites in 18 countries worldwide across Europe, North America, South America, Asia, and Australia. Patients with mPDAC and Eastern Cooperative Oncology Group performance status score 0 or 1 were randomly assigned (1:1) to receive NALIRIFOX (liposomal irinotecan 50 mg/m2, oxaliplatin 60 mg/m2, leucovorin 400 mg/m2, and fluorouracil 2400 mg/m2, administered sequentially as a continuous intravenous infusion over 46 h) on days 1 and 15 of a 28-day cycle or nab-paclitaxel 125 mg/m2 and gemcitabine 1000 mg/m2, administered intravenously, on days 1, 8, and 15 of a 28-day cycle. Balanced block randomisation was stratified by geographical region, performance status, and liver metastases, managed through an interactive web response system. The primary endpoint was overall survival in the intention-to-treat population, evaluated when at least 543 events were observed across the two treatment groups. Safety was evaluated in all patients who received at least one dose of study treatment. This completed trial is registered with ClinicalTrials.gov, NCT04083235. Findings Between Feb 19, 2020 and Aug 17, 2021, 770 patients were randomly assigned (NALIRIFOX, 383; nab-paclitaxel–gemcitabine, 387; median follow-up 16·1 months [IQR 13·4–19·1]). Median overall survival was 11·1 months (95% CI 10·0–12·1) with NALIRIFOX versus 9·2 months (8·3–10·6) with nab-paclitaxel–gemcitabine (hazard ratio 0·83; 95% CI 0·70–0·99; p=0·036). Grade 3 or higher treatment-emergent adverse events occurred in 322 (87%) of 370 patients receiving NALIRIFOX and 326 (86%) of 379 patients receiving nab-paclitaxel–gemcitabine; treatment-related deaths occurred in six (2%) patients in the NALIRIFOX group and eight (2%) patients in the nab-paclitaxel–gemcitabine group. Interpretation Our findings support use of the NALIRIFOX regimen as a possible reference regimen for first-line treatment of mPDAC.Ipsen

Pancreatic ductal adenocarcinoma cell lines display a plastic ability to bi‑directionally convert into cancer stem cells

Pancreatic ductal adenocarcinoma (PDAC) is often diagnosed when metastatic events have occurred. Cancer stem cells (CSCs) play an important role in tumor initiation, metastasis, chemoresistance and relapse. A growing number of studies have suggested that CSCs exist in a dynamic equilibrium with more differentiated cancer cells via a bi‑directional regeneration that is dependent on the environmental stimuli. In this investigation, we obtain, by using a selective medium, PDAC CSCs from five out of nine PDAC cell lines, endowed with different tumorsphere‑forming ability. PDAC CSCs were generally more resistant to the action of five anticancer drugs than parental cell lines and were characterized by an increased expression of EpCAM and CD44v6, typical stem cell surface markers, and a decreased expression of E‑cadherin, the main marker of the epithelial state. PDAC CSCs were able to re‑differentiate into parental cells once cultured in parental growth condition, as demonstrated by re‑acquisition of the epithelial morphology, the decreased expression levels of EpCAM and CD44v6 and the increased sensitivity to anticancer drugs. Finally, PDAC CSCs injected into nude mice developed a larger subcutaneous tumor mass and showed a higher metastatic activity compared to parental cells. The present study demonstrates the ability to obtain CSCs from several PDAC cell lines and that these cells are differentially resistant to various anticancer agents. This variability renders them a model of great importance to deeply understand pancreatic adenocarcinoma biology, to discover new biomarkers and to screen new therapeutic compounds

Quality of life in metastatic pancreatic cancer patients receiving liposomal irinotecan plus 5-fluorouracil and leucovorin

Abstract Background The NAPOLI-1 study (NCT01494506) reported that liposomal irinotecan plus 5-fluorouracil and leucovorin (nal-IRI+5-FU/LV) improved overall survival vs 5-FU/LV with manageable toxicity in patients with metastatic pancreatic adenocarcinoma previously treated with gemcitabine-based therapy. Yet, clinicians need treatment strategies that also maintain the patient's health-related quality of life (HRQOL). Here, we report the HRQOL data. Methods Patients completed the European Organisation for Research and Treatment of Cancer QOL core questionnaire C30 (EORTC QLQ-C30) at baseline, every 6 weeks, and at 30 days after discontinuation of study treatment. Patient-reported outcomes (PROs) were scored according to EORTC guidelines. nal-IRI+5-FU/LV HRQOL was compared with 5-FU/LV. The PRO population comprised intent-to-treat patients who completed baseline and at least one subsequent assessment on the EORTC QLQ-C30. Data were also analysed for missingness. Results Of 236 patients in the intent-to-treat population, 128 (54.2%) comprised the PRO population (71 in the nal-IRI+5-FU/LV arm; 57 the in 5-FU/LV arm). Of the remaining 108 patients (45.8%) not included in the PRO population, most progressed rapidly, making participation difficult. Median change from baseline was ≤10 points at weeks 6 and 12 in global health status or functional and symptom scale scores, except for fatigue, which deteriorated by 11.1 points with nal-IRI+5-FU/LV but did not change vs 5-FU/LV. The proportion of patients whose HRQOL improved or deteriorated was not significantly different between the arms. Conclusion In the NAPOLI-1 study, HRQOL was maintained with nal-IRI+5-FU/LV in patients with metastatic pancreatic adenocarcinoma previously treated with a gemcitabine-based regimen, while survival was significantly extended

Molecular analysis of a male breast cancer patient with prolonged stable disease under mTOR/PI3K inhibitors BEZ235/everolimus

The mTORC1 inhibitor everolimus (Afinitor/RAD001) has been approved for multiple cancer indications, including ER(+)/HER2(-) metastatic breast cancer. However, the combination of everolimus with the dual PI3K/mTOR inhibitor BEZ235 was shown to be more efficacious than either everolimus or BEZ235 alone in preclinical models. Herein, we describe a male breast cancer (MBC) patient who was diagnosed with hormone receptor-positive (HR(+))/HER2(-) stage IIIA invasive ductal carcinoma and sequentially treated with chemoradiotherapy and hormonal therapy. Upon the development of metastases, the patient began a 200 mg twice-daily BEZ235 and 2.5 mg weekly everolimus combination regimen. The patient sustained a prolonged stable disease of 18 mo while undergoing the therapy, before his tumor progressed again. Therefore, we sought to both better understand MBC and investigate the underlying molecular mechanisms of the patient's sensitivity and subsequent resistance to the BEZ235/everolimus combination therapy. Genomic and immunohistochemical analyses were performed on samples collected from the initial invasive ductal carcinoma pretreatment and a metastasis postprogression on the BEZ235/everolimus combination treatment. Both tumors were relatively quiet genomically with no overlap to recurrent MBC alterations in the literature. Markers of PI3K/mTOR pathway hyperactivation were not identified in the pretreatment sample, which complements previous reports of HR(+) female breast cancers being responsive to mTOR inhibition without this activation. The postprogression sample, however, demonstrated greater than fivefold increased estrogen receptor and pathogenesis-related protein expression, which could have constrained the PI3K/mTOR pathway inhibition by BEZ235/everolimus. Overall, these analyses have augmented the limited episteme on MBC genetics and treatment

Induction of immunosuppressive functions and NF-\u3baB by FLIP in monocytes

Immunosuppression is a hallmark of tumor progression, and treatments that inhibit or deplete monocytic myeloid-derived suppressive cells could promote anti-tumor immunity. c-FLIP is a central regulator of caspase-8-mediated apoptosis and necroptosis. Here we show that low-dose cytotoxic chemotherapy agents cause apoptosis linked to c-FLIP down-regulation selectively in monocytes. Enforced expression of c-FLIP or viral FLIP rescues monocytes from cytotoxicity and concurrently induces potent immunosuppressive activity, in T cell cultures and in vivo models of tumor progression and immunotherapy. FLIP-transduced human blood monocytes can suppress graft versus host disease. Neither expression of FLIP in granulocytes nor expression of other anti-apoptotic genes in monocytes conferred immunosuppression, suggesting that FLIP effects on immunosuppression are specific to monocytic lineage and distinct from death inhibition. Mechanistically, FLIP controls a broad transcriptional program, partially by NF-\u3baB activation. Therefore, modulation of FLIP in monocytes offers a means to elicit or block immunosuppressive myeloid cells

A Case Matched Gender Comparison Transcriptomic Screen Identifies eIF4E and eIF5 as Potential Prognostic and Tractable Biomarkers in Male Breast Cancer

Purpose: Breast cancer (BC) affects both genders, but is understudied in men. Although still rare, male BC is being diagnosed more frequently. Treatments are wholly informed by clinical studies conducted in women, based on assumptions that underlying biology is similar. Experimental design: A transcriptomic investigation of male and female BC was performed, confirming transcriptomic data in silico. Biomarkers were immunohistochemically assessed in 697 MBCs (n=477, training; n=220, validation set) and quantified in pre- and post-treatment samples from a male BC patient receiving Everolimus and PI3K/mTOR inhibitor. Results: Gender-specific gene expression patterns were identified. eIF transcripts were up-regulated in MBC. eIF4E and eIF5 were negatively prognostic for overall survival alone (Log rank; p=0.013; HR=1.77, 1.12-2.8 and p=0.035; HR=1.68, 1.03-2.74, respectively), or when co-expressed (p=0.01; HR=2.66, 1.26-5.63), confirmed in the validation set. This remained upon multivariate Cox regression analysis (eIF4E p=0.016; HR 2.38 (1.18-4.8), eIF5 p=0.022; HR 2.55 (1.14-5.7); co-expression p=0.001; HR=7.04 (2.22-22.26)). Marked reduction in eIF4E and eIF5 expression was seen post BEZ235/Everolimus, with extended survival. Conclusions: Translational initiation pathway inhibition could be of clinical utility in male BC patients overexpressing eIF4E and eIF5. With mTOR inhibitors which target this pathway now in the clinic, these biomarkers may represent new targets for therapeutic intervention, although further independent validation is required

Smad- dependent and Smad- independent pathways in TGF-β signaling as therapeutic targets in pancreatic cancer

Pancreatic cancer (PC) remains one of the most lethal and poorly understood human malignancies. Most PC patients present with inoperable disease, or develop metastases after surgery, and conventional therapies are usually ineffective in treating this disease. It is evident that novel therapies remain to be developed. Transforming Growth Factor β (TGF-β) is a family of dimeric olypeptide growth factors that initiate cell signaling by dimerizing the TGF-β type I (TβRI) and type II (TβRII) serine/threonine kinase receptors, which, in turn, propagate the signal by activating downstream Smad-dependent and Smad-independent pathways. TGF-β signaling plays a key role in the tumor progression and metastasis of many different types of tumor cells. The overexpression of TGF-β has been reported in most tumor types. In human PC, increased levels of TGF-β have been significantly associated with venous invasion, advanced tumor stages, progressive disease, shorter patient survival duration, and liver metastases. The effect of TGF-β is not limited to the tumor cells themselves in vivo. It has been recently highlighted its paracrine mechanism in the interactions between various cell populations in the tumor microenvironment. Hence its pleiotropic role in PC biology, the TGF-β Smad- dependent and independent pathways represent a critical area for the development of novel strategies for the treatment of PC patients. We initially hypothesized that targeting the Smad- dependent pathways by inhibiting TRI/II kinase activity with the novel inhibitor LY2109761 would suppress PC growth and metastatic processes. The effect of LY2109761 was evaluated on soft-agar growth, migration, invasion using a fibroblast coculture model, and anoikis by annexin V flow cytometric analysis. The efficacy of LY2109761 on tumor growth, survival and reduction of spontaneous metastasis was evaluated in an orthotopic murine model of metastatic PC expressing both luciferase and green fluorescence proteins (L3.6pl/GLT). To determine whether PC cells or the cells in the liver microenvironment were involved in LY2109761-mediated reduction of liver metastasis, we used a model of experimental liver metastasis. LY2109761 significantly inhibited the L3.6pl/GLT soft-agar growth; suppressed both basal and TGF-induced cell migration and invasion, and induced anoikis. In vivo, LY2109761 in combination with gemcitabine (GEM) significantly reduced the tumor burden, prolonged the survival, and reduced spontaneous abdominal metastases. Results from the experimental liver metastasis models indicate an important role for targeting TRI/II kinase activity on tumor and liver microenvironment cells in suppressing liver metastasis. Resistance to chemotherapeutic drugs poses one of the greatest challenges in PC research. The Smad-independent effector TGF-- Activated Kinase- 1 (TAK-1) is a MAP3K essential for the activation of NFB and AP1 transcription factors. cIAP2 is a NFB and AP1 target gene that regulates apoptosis by direct inhibition of caspases. We hypothesized that TAK1 is responsible for PC chemoresistance by regulating NFB- and AP-1- mediated transcription of cIAP2. The expression of TAK-1 in PC cell lines was studied by Western blot. TAK-1 expression was silenced by shRNA in AsPC-1, Panc-1, and Panc-28 cell lines. NFB and AP-1 activation was analyzed by EMSA. Apoptosis was quantified using cleavage of caspase-3 and PARP1 and DNA fragmentation. MTT assays were used to assess the in vitro chemopotentiation of GEM, oxaliplatin (OX), and SN-38. TAK-1 kinase activity was targeted using an orally available small molecule selective inhibitor (Ki in enzymatic assay: TAK1 = 13 nM; p38 > 20 μM; IKK > 20 μM) provided by Eli Lilly Research Labs. In vivo activity of the TAK-1 inhibitor alone and in combination with GEM was evaluated in an orthotopic nude mouse model with luciferase-expressing AsPC-1 PC cells. TAK-1 protein was overexpressed in all PC cell lines studied but not in normal pancreatic ductal epithelial cells. shRNA knockdown of TAK-1 completely suppressed both NFB and AP1 DNA binding activities. As a result, cIAP2 expression was completely suppressed, inducing a proapoptotic phenotype as demonstrated by higher levels of cleaved caspase-3 and PARP-1 and by DNA fragmentation. shRNA silencing of TAK-1 in AsPC-1, Panc-1, and Panc-28 cell lines resulted in significantly higher in vitro sensitivity to GEM, OX, and SN-38, compared to the respective control cell lines. In vitro, the TAK1 inhibitor alone demonstrated potent cytotoxic activity (IC50 5-39 nM) and suppressed NFB DNA binding activity in all three PC cell lines studied. In combination, the TAK1 inhibitor strongly potentiated the cytotoxic activities of GEM, OX, or SN-38 in all three PC cell lines. In nude mice, oral administration of the TAK-1 inhibitor plus GEM significantly reduced tumor burden and prolonged mice survival. In our study we demonstrated that targeting the Smad- dependent pathways by inhibiting TRI/II kinase activity on PC cells or the cells of the liver microenvironment represents a novel therapeutic approach to prevent PC metastasis. Our study is the first to demonstrate that the genetic silencing or inhibition of the Smad-independent effector TAK1 is a valid approach to revert in vivo the intrinsic chemoresistance of PC. Specifically designed clinical trials need to be conducted to evaluate the therapeutic effect of LY2109761 as a neoadjuvant treatment in patients with resectable PC. The TAK-1 inhibitor used in this study is an exciting drug that warrants further development for the treatment of advanced PC