Agent Of Several Cancers Research Articles

Rationale and design of the Self-TI Study protocol: a cross-sectional human papillomavirus self-testing pilot study among transgender adults in England

IntroductionPersistent infection with high-risk human papillomavirus (HPV) is the causal agent of several cancers including cervical, anal and oropharyngeal cancer. Transgender men and transmasculine non-binary (TMNB) people with a cervix...

Dysregulated MicroRNAs Contribute to Chemotherapy-Resistance of Pleural Mesothelioma

Abstract Background Dysregulated expression of microRNAs has been shown to contribute to response to chemotherapeutic agents in several cancers, including pleural mesothelioma (PM). In previous work, we found expression of several tissue-microRNAs to correlate with patient’s response to cisplatin-pemetrexed. Aims We want to understand how altering the expression of these microRNAs affects cell growth and chemo response. Methods PM cell lines MSTO-211H (biphasic), H28, Meso-1, Mero-82 (epithelioid) and non-malignant MeT-5A were reverse transfected with synthetic microRNA mimics for 15 candidates. Following transfection, cell growth, colony forming ability, and migratory potential were assessed using standard assays. Furthermore, transfected cells were exposed to increasing concentrations of cisplatin to evaluate sensitivity to these drugs. Results Overexpression of seven candidate microRNAs resulted in growth inhibition in all investigated PM cell lines five days post transfection. The strongest effects were observed for miR-380-5p, miR-221-3p, miR-210, miR-625-3p, and miR-19b, which reduced cell growth to 30-60%. Growth of non-malignant MeT-5A cells remained largely unaffected (Fig. A). Overexpression of those microRNAs also resulted in a strong reduction of colony forming ability (Fig. B), while an effect on wound healing capacity (migration) could not be observed. Finally, we found that especially overexpression of miR-221-3p, miR-625-3p and miR-19b was able to sensitise cells towards cisplatin, with the strongest effect observed in MSTO-211H for miR-221-3p (IC50 from 17.6µM to 2.7µM) and miR-19b (IC50 from 17.6µM to 0.7µM), and in Mero-82 for miR-625-3p (IC50 from 3.75µM to 0.8µM) (Fig. C). Conclusion We show that overexpression of several microRNAs has the potential to alter PM cell growth and colony forming ability. Furthermore, microRNA overexpression can sensitise cells towards cisplatin, although the degree of sensitisation varies between different cell lines. Current analyses focus on the response towards the cisplatin/pemetrexed doublet and on the effect on cell cycle and expression of associated genes.

CHARLI: A phase Ib/II trial of ipilimumab-nivolumab-denosumab or nivolumab-denosumab in patients with unresectable stage III and IV melanoma.

9525 Background: Denosumab (deno) is an antibody directed against Receptor Activator of NF Kappa-b ligand (RANKL) with established indications as a bone anti-resorptive agent in several cancers. Pre-clinical studies and several case series suggest anti-RANKL can enhance the anti-tumor effect of immune checkpoint inhibitors possibly via modulation of Treg and M2 macrophages. We did a multicentre phase Ib/II trial (NCT03161756) to investigate the safety and efficacy of deno in combination with nivolumab (nivo) or ipilimumab-nivolumab (ipi-nivo). Methods: Patients (pts) with unresectable stage III or IV melanoma were recruited to either Arm A (nivo-deno) or Arm B (ipi-nivo-deno) as first-line therapy. In Arm A pts received nivo 3 mg/kg IV q2 weekly for 4 doses and deno 120 mg SC on D1, D8, D15, D29 and then maintenance nivo 480 mg IV with deno 120 mg SC q 4 weekly. In Arm B pts received combined ipi 3mg/kg with nivo 1 mg/kg IV 3 weekly for 4 doses with deno 120 mg SC on D1, D8, D15, D29 followed by 4 weekly maintenance nivo 480 mg with deno 120 mg. Co-primary endpoints were median PFS and grade 3-4 treatment related adverse events (TRAE) of interest. Secondary endpoints were objective response rate (ORR) and overall survival (OS). Results: 27 pts (15 males, median age 67 years, 48% (13/27) stage IVM1c, 26% elevated LDH, 31% BRAFV600 mutant) were enrolled in Arm A and 24 of 25 evaluable pts (16 males, median age 62, 46% (11/24) stage IVM1c, 13% (3/24) stage IVM1d, 30% elevated LDH and 33% BRAFV600 mutant) were enrolled in Arm B. Median follow up was 30.8 months (m) for Arm A and 24.8 m for Arm B. The RECIST 1.1 ORR was 56% (n=15/27, 15% [4/27] complete responses [CR] and 41% [11/27] partial response [PR]) for Arm A and 71% (17/24, 25% [6/24) CR and 46% [11/24] PR) for Arm B. The median PFS in both arms has not been reached with 12 month PFS rates of 59% (95%CI: 43-81) and 63% (95%CI: 46-88) for Arms A & B, respectively. Grade 3-4 TRAE were 11% (3/27) in Arm A and 71% (17/24) in Arm B. Common TRAE (≥10%) in Arm A was rash (52%), pruritus (30%), fatigue (26%), nausea (19%), diarrhoea/colitis (15%), arthralgia (11%), vitiligo (11%), hyperthyroidism (11%). Common TRAE (≥10%) in Arm B included rash (63%), fatigue (54%), diarrhoea/colitis (54%), hepatitis (46%), hyperthyroidism (29%), pneumonitis (29%), pruritus (25%), increased GGT (17%), hypothyroidism (17%), hypocalcaemia (13%). Arm B G3-4 TRAE included diarrhoea/colitis (25%), pneumonitis (17%) and hepatitis (13%). Conclusions: Nivo-deno and ipi-nivo-deno had numerically similar G3-4 TRAE compared to nivo and ipi-nivo. The median PFS, 12 month PFS and ORR for nivo-deno and ipi-nivo-deno are encouraging compared with CHECKMATE 067. Clinical trial information: NCT03161756 . [Table: see text]

Metformin and simvastatin exert additive antitumour effects in glioblastoma via senescence-state: clinical and translational evidence.

Glioblastoma is one of the most devastating and incurable cancers due to its aggressive behaviour and lack of available therapies, being its overall-survival from diagnosis ∼14-months. Thus, identification of new therapeutic tools is urgently needed. Interestingly, metabolism-related drugs (e.g., metformin/statins) are emerging as efficient antitumour agents for several cancers. Herein, we evaluated the invitro/invivo effects of metformin and/or statins on key clinical/functional/molecular/signalling parameters in glioblastoma patients/cells. An exploratory-observational-randomized retrospective glioblastoma patient cohort (n=85), human glioblastoma/non-tumour brain human cells (cell lines/patient-derived cell cultures), mouse astrocytes progenitor cell cultures, and a preclinical xenograft glioblastoma mouse model were used to measure key functional parameters, signalling-pathways and/or antitumour progression in response to metformin and/or simvastatin. Metformin and simvastatin exerted strong antitumour actions in glioblastoma cell cultures (i.e., proliferation/migration/tumoursphere/colony-formation/VEGF-secretion inhibition and apoptosis/senescence induction). Notably, their combination additively altered these functional parameters vs. individual treatments. These actions were mediated by the modulation of key oncogenic signalling-pathways (i.e., AKT/JAK-STAT/NF-κB/TGFβ-pathways). Interestingly, an enrichment analysis uncovered a TGFβ-pathway activation, together with AKT inactivation, in response to metformin+simvastatin combination, which might be linked to an induction of the senescence-state, the associated secretory-phenotype, and to the dysregulation of spliceosome components. Remarkably, the antitumour actions of metformin+simvastatin combination were also observed invivo [i.e., association with longer overall-survival in human, and reduction in tumour-progression in a mouse model (reduced tumour-size/weight/mitosis-number, and increased apoptosis)]. Altogether, metformin and simvastatin reduce aggressiveness features in glioblastomas, being this effect significantly more effective (invitro/invivo) when both drugs are combined, offering a clinically relevant opportunity that should be tested for their use in humans. Spanish Ministry of Science, Innovation and Universities; Junta de Andalucía; CIBERobn (CIBER is an initiative of Instituto de Salud Carlos III, Spanish Ministry of Health, Social Services and Equality).

Niclosamide causes lysosome-dependent cell death in endometrial cancer cells and tumors.

Endometrial cancer is the most common female cancer showing continuous rise in its incidence and mortality rate. Despite the extensive research efforts in cancer therapeutics, still there is a lack of effective treatment options and the outcome is poor for patients with advanced and recurrent endometrial cancers. In this study, we aimed to evaluate the efficacy of niclosamide (NIC) against endometrial cancer. NIC is an FDA-approved anti-helminthic drug, which has been recently extensively studied as a potent anti-cancerous agent in several cancers. The anti-cancerous activity of NIC was analyzed in-vitro (ANC3A, Hec1B, and Ishikawa endometrial cancer cell lines) by cell viability-, soft agar-, invasion- and migration- assay. The action mechanism of NIC was demonstrated by western blot analysis and immune-fluorescence imaging and validated by specific inhibitors. The in-vivo efficacy of NIC was studied in the Ishikawa xenograft animal model. NIC effectively suppressed the viability (IC50<1 μM), colony formation ability, migration, and invasion of all endometrial cancer cells tested. We demonstrated that NIC inhibited AKT/mTOR signaling pathway and induced apoptosis and autophagy in endometrial cancer cells. Further study demonstrated that although NIC induced autophagosome formation, it inhibits autolysosome formation. In addition, we observed that NIC induced BAX co-localization with lysosome and inhibited Cathepsin B maturation from pro-cathepsin B, thereby inducing the lysosomal membrane permeability and release of hydrolytic enzymes from the lysosome to cytosol, which eventually contributed cell death. NIC also inhibited tumor weight and volume in the Ishikawa xenograft animal model without having any evidence of toxicity. Due to its potent anti-cancerous activity and safety profile, NIC seems to be a promising agent for human endometrial cancer therapeutics.

Salvianolic Acid B Suppresses Non-Small-Cell Lung Cancer Metastasis through PKM2-Independent Metabolic Reprogramming.

Objective Salvianolic acid B (Sal B) has been demonstrated to be a potential chemoprevention agent for several cancers. Herein, we investigated the pharmacological function of Sal B on non-small-cell lung cancer (NSCLC) metastasis. Methods Two NSCLC cell lines (NCI-H2030 and NCI-H1650) were disposed of by 200 μM Sal B or 10 μM PKM2 agonist TEPP-46. Wound healing and transwell experiments were implemented for analyzing migratory and invasive capacities. Epithelial-to-mesenchymal transition (EMT) markers β-catenin and E-cadherin were measured via western blotting. Cellular bioenergetics were evaluated with glucose uptake, lactate production, enolase activity, cellular ATP levels, as well as seahorse-based oxygen consumption rate (OCR), extracellular acidification rate (ECAR) analysis. Metabolic reprogramming markers PKM2, LDHA, and GLUT1 were detected via western blotting and immunofluorescence. Results The results showed that Sal B disposal weakened the migration and invasion of NCI-H2030 and NCI-H1650 cells and inactivated the EMT process according to downregulation of β-catenin and upregulation of E-cadherin. Sal B-treated NSCLC cells displayed decreased glucose uptake, lactate production, enolase activity, cellular ATP levels, OCR, and ECAR, indicating a reduction in metabolic reprogramming. Additionally, Sal B downregulated the expression of PKM2, LDHA, and GLUT1. TEPP-46 may reverse the inhibitory effect of Sal B on metastasis as well as metabolic reprogramming. Conclusion Our findings provide evidence that Sal B enables to weaken NSCLC metastasis through PKM2-independent metabolic reprogramming, which sheds light on the promising therapeutic usage of Sal B in treating NSCLC.

Combination of berberine and low glucose inhibits gastric cancer through the PP2A/GSK3β/MCL-1 signaling pathway

Intermittent fasting as an adjuvant therapy in clinical practice is an emerging treatment modality to target tumor growth by reducing glucose utilization. Berberine, an alkaloid extracted from the traditional Chinese medicine Coptidis Rhizoma, has been shown to be a safe and effective antitumor agent in several cancers. Hence, the purpose of the present study was to investigate the effects of the combination of berberine and low glucose on gastric cancer. Our results showed that the combination of berberine and low glucose effectively inhibited cell viability, promoted apoptosis, and reduced the migration ability of MGC803 cells. In addition, the combination was shown to activate the PP2A/GSK3β signaling axis, leading to the downregulation of the downstream pro-survival protein MCL-1, which leads to the death of gastric cancer cells. In addition, the inhibitor of GSK3β partially reversed the effect of this combination on MGC803 cells. In vivo experiments demonstrated that berberine effectively impaired the growth of xenograft tumors, when administered during intermittent fasting (hypoglycemic conditions), and was well tolerated by nude mice without the occurrence of any adverse effects. Based on these results, we conclude that the berberine/low-glucose combination can inhibit the growth of gastric cancer through the PP2A/GSK3β/MCL-1 signaling pathway. Accordingly, this combination of drugs and lifestyle may become a new type of safe and effective anti-cancer therapy.

The Role of Bacteria in KSHV Infection and KSHV-Induced Cancers.

Simple SummaryThe aim of this article is to review the complex interactions of bacteria with Kaposi’s sarcoma-associated herpesvirus (KSHV) infection and KSHV-induced cancers. KSHV is causally associated with multiple cancers including Kaposi’s sarcoma (KS) and primary effusion lymphoma. Among patients coinfected by HIV and KSHV, patients with KS have a distinct oral microbiome compared to patients without KS. Moreover, KSHV patients have increased levels of salivary bacterial pathogen-associated molecular patterns compared to KSHV-negative patients. KSHV-associated bacterial species can increase KSHV replication and dissemination, and enhance cell proliferation of KSHV-transformed cells. The analysis of bacterial biomarkers associated with KSHV may help improve our understanding of the mechanisms driving KSHV-induced oncogenesis and identify novel targets for improving therapies of KSHV-related cancers.The objective of this article is to review the current status of the bacteria-virus interplay in Kaposi’s sarcoma-associated herpesvirus (KSHV) infection and KSHV-driven cancers. KSHV is the etiological agent of several cancers, including Kaposi’s sarcoma (KS) and primary effusion lymphoma. Due to immunosuppression, patients with KSHV are at an increased risk for bacterial infections. Moreover, among patients coinfected by HIV and KSHV, patients with KS have distinct oral microbiota compared to non-KS patients. Bacterial biomarkers associated with KSHV-driven cancers can provide insights in discerning the mechanisms of KSHV-induced oncogenesis. For example, pathogen-associated molecular patterns and bacterial products of certain bacterial species can regulate the expression of KSHV lytic and latent genes, thereby affecting viral replication and dissemination. In addition, infection with distinct opportunistic bacterial species have been associated with increased cell proliferation and tumorigenesis in KSHV-induced cancers through activation of pro-survival and -mitogenic cell signaling pathways. By elucidating the various mechanisms in which bacteria affect KSHV-associated pathogenesis, we will be able to pinpoint therapeutic targets for KSHV infection and KSHV-related cancers.

Paracetamol Intake and Hematologic Malignancies: A Meta-Analysis of Observational Studies.

Hematologic malignancies cause more than half a million deaths every year worldwide. Analgesics were suggested as chemopreventive agents for several cancers but so far, results from individual studies about the relationship between paracetamol (acetaminophen) use and hematologic malignancies are conflicting. Therefore, we decided to perform a systematic review and meta-analysis. We retrieved studies published in any language by systematically searching Medline, Embase, Conference Proceedings Citation Index, Open Access Theses and Dissertations, and the five regional bibliographic databases of the World Health Organization until December 2020. Pooled odds ratios (OR) and their 95% confidence intervals (CI) were calculated according to the inverse of their variances. We performed separate analyses by histologic type. We also evaluated publication bias and assessed quality. A total of 17 study units met our inclusion criteria. The results show an association of hematologic malignancies with any paracetamol intake (OR 1.49, 95% CI 1.23–1.80) and with high paracetamol intake (OR 1.77, 95% CI 1.45–2.16). By subtype, risk was higher for multiple myeloma (OR 2.13, 95% CI 1.54–2.94) for any use and OR 3.16, 95% CI 1.96–5.10 for high intake, while risk was lower and non-significant for non-Hodgkin lymphoma. This meta-analysis provides evidence that paracetamol intake may be associated with hematologic malignancies and suggests that a dose–response effect is plausible. These results are unlikely to be due to publication bias or low quality of studies. Future research should focus on assessing the dose–response relationship.

Effects of taxol on neuronal differentiation of postnatal neural stem cells cultured from mouse subventricular zone

Taxol (paclitaxel), a chemotherapeutic agent for several cancers, can adversely affect the peripheral nervous system. Recently, its negative impact on cognitive function in cancer patients has become evident. In rodents, taxol impaired learning and memory, with other possible negative effects on the brain. In this study, we investigated the effects of taxol on cultured neural stem cells (NSCs) from the mouse neurogenic region, the subventricular zone (SVZ). Taxol significantly decreased both proliferation and neuronal differentiation of NSCs. Transient treatment with taxol for one day during a 4-day differentiation greatly decreased neurogenesis along with an abnormal cell cycle progression. Yet, taxol did not kill differentiated Tuj1+ neurons and those neurons had longer neurites than neurons under control conditions. For glial differentiation, taxol significantly reduced oligodendrogenesis as observed by immunostaining for Olig2 and O4. However, differentiation of astrocytes was not affected by taxol. In contrast, differentiated oligodendrocytes were extremely sensitive to taxol. Almost no Olig2-positive cells were observed after three days of treatment with taxol. Taxol has distinct effects on neurons and glial cells during their production through differentiation from NSCs as well as post-differentiation. Thus, we suggest that taxol might interfere with neurogenesis of NSCs possibly through a disturbance in the cell cycle and may eliminate differentiated oligodendrocytes.

Mesenchymal Stem Cells: A New Generation of Therapeutic Agents as Vehicles in Gene Therapy.

In recent years, mesenchymal stem cells (MSCs) as a new tool for therapeutic gene delivery in clinics have attracted much attention. Their advantages cover longer lifespan, better isolation, and higher transfection efficiency and proliferation rate. MSCs are the preferred approach for cell-based therapies because of their in vitro self-renewal capacity, migrating especially to tumor tissues, as well as anti-inflammatory and immunomodulatory properties. Therefore, they have considerable efficiency in genetic engineering for future clinical applications in cancer gene therapy and other diseases. For improving therapeutic efficiency, targeted therapy of cancers can be achieved through the sustained release of therapeutic agents and functional gene expression induction to the intended tissues. The development of a new vector in gene therapy can improve the durability of a transgene expression. Also, the safety of the vector, if administered systemically, may resolve several problems, such as durability of expression and the host immune response. Currently, MSCs are prominent candidates as cell vehicles for both preclinical and clinical trials due to the secretion of therapeutic agents in several cancers. In the present study, we discuss the status of gene therapy in both viral and non-viral vectors along with their limitations. Throughout this study, the use of several nano-carriers for gene therapy is also investigated. Finally, we critically discuss the promising advantages of MSCs in targeted gene delivery, tumor inhibition and their utilization as the gene carriers in clinical situations.

Homologous Recombination Repair Truncations Predict Hypermutation in Microsatellite Stable Colorectal and Endometrial Tumors.

INTRODUCTION:Somatic mutations in BRCA1/2 and other homologous recombination repair (HRR) genes have been associated with sensitivity to PARP inhibitors and/or platinum agents in several cancers, whereas hypermutant tumors caused by alterations in POLE or mismatch repair genes have demonstrated robust responses to immunotherapy. We investigated the relationship between somatic truncations in HRR genes and hypermutation in colorectal cancer (CRC) and endometrial cancer (EC).METHODS:We analyzed the mutational spectra associated with somatic BRCA1/2 truncations in multiple genomic cohorts (N = 2,335). From these results, we devised a classifier incorporating HRR genes to predict hypermutator status among microsatellite stable (MSS) tumors. Using additional genomic cohorts (N = 1,439) and functional in vivo assays, we tested the classifier to disambiguate POLE variants of unknown significance and identify MSS hypermutators without somatic POLE exonuclease domain mutations.RESULTS:Hypermutator phenotypes were prevalent among CRCs with somatic BRCA1/2 truncations (50/62, 80.6%) and ECs with such mutations (44/47, 93.6%). The classifier predicted MSS hypermutators with a cumulative true-positive rate of 100% in CRC and 98.0% in EC and a false-positive rate of 0.07% and 0.63%. Validated by signature analyses of tumor exomes and in vivo assays, the classifier accurately reassigned multiple POLE variants of unknown significance as pathogenic and identified MSS hypermutant samples without POLE exonuclease domain mutations.DISCUSSION:Somatic truncations in HRR can accurately fingerprint MSS hypermutators with or without known pathogenic exonuclease domain mutations in POLE and may serve as a low-cost biomarker for immunotherapy decisions in MSS CRC and EC.

Prevalence of Major Cardiac Events of Anthracycline-Induced Cardiotoxicity in Southwestern Iran: Different Response Patterns to Cumulative Dose

Background:Anthracyclines are widely used chemotherapeutic agents in several cancers. Since its use, survival improved significantly among cancer patients and has been reported to be up to 80%. However, anthracyclines possess several cardiac, renal and hematological toxicities which limit their use in practice. Cardiotoxicity is still the most important and dose-limiting side effect of anthracycline treatment. Here we aimed to investigate the frequency of anthracyclineinduced cardiomyopathy in pediatric malignancies in Khuzestan Province, Iran.Methods:A total of 112 patients were enrolled in the present study. Patients were allocated to the case or control group based on receiving anthracycline. Echocardiographic examinations were performed by a cardiologist. Electrocardiograms were also recorded.Results:We showed that cancer patients who underwent anthracycline treatment showed cardiomyopathy as defined by lower LVEF (Left Ventricular Ejection Fraction) among patients (p = 0.041). Abnormal LVEF was reported with a frequency of about 9.5% in patients (p = 0.026). However, LVFS (Left Ventricular Fraction Shortening), QRS voltage and QT interval did not differ significantly between treatment and control groups. Our data analysis revealed that this difference is mainly related to high cumulative dose since high cumulative dose of anthracycline (&gt;300 mg/m2) leads to lower LVEF and LVFS and higher QRS voltage in comparison with lower cumulative dose (&lt;300 mg/m2) and control group; but there was no significant difference between low dose and control group. Different age groups and type of malignancy including hematological and solid tumors did not show any significant differences for echocardiographic and electrocardiograms parameters.Conclusion:In our study, lower LVEF among patients who received anthracyclines were mainly related to a high cumulative dose of anthracyclines, which emphasizes the effect of cumulative dose for cardiotoxic effects. Larger studies are needed to investigate possible other risk factors for cardiotoxicity.

668TiP - Phase II study of avelumab in combination with cetuximab as a rechallenge strategy in pre-treated RAS wild type metastatic colorectal cancer patients: CAVE (cetuximab-avelumab) colon

BackgroundThe immune system plays a pivotal role in modulating response to monoclonal antibodies therapy in cancer. Novel immune-checkpoint inhibitors have demonstrated potent efficacy alone and in combinations with cytotoxic agents in several cancers. In this regard, avelumab in combination with cetuximab might be a relevant re-challenge strategy in RAS wild type (WT) metastatic colorectal cancer (mCRC) patients treated in first line with chemotherapy (CT) in combination with anti-epidermal growth factor receptor (EGFR) drugs and who achieved a complete or partial response. Trial designCAVE Colon is a single arm, multi-center phase II study designed to evaluate the efficacy of avelumab and cetuximab in pre-treated RAS WT mCRC patients. Eligible patients: pathologically confirmed KRAS (exon 2, 3 and 4) and NRAS (exon 2, 3 and 4) WT mCRC treated with a first line CT in combination with an anti-EGFR agent with a major response achieved (complete or partial), who have progressed to a second line therapy, and received no prior immunotherapy. Primary endpoint is overall survival; secondary endpoints are overall response rate according to RECIST 1.1, progression free survival and safety profile. This study seeks to demonstrate a median OS of 11 months by the experimental combination for comparison with historical median OS 8.0 with standard third line treatments, which correspond to an improvement of OS at 6 months from 40% to 57%. With 80% power and 1-sided 5% level test, it was estimated that it would be needed to enrol 66 patients. Considering a potential dropout of approximately 15% of patients, a total of 75 patients will be recruited. The accrual period, started in August 2018, will be 18 months and the total duration of the study will be 36 months. As per April 22nd 2019, thirty-five patients have been enrolled and started the treatment with avelumab 10mg/kg q14 as a 1-hour i.v. infusion and cetuximab at 400mg/m2 over 2-hour and subsequently 250mg/m2 q7 as 1-hour i.v. infusion until disease progression or unacceptable toxicity. Clinical trial identification2017-004392-32. Legal entity responsible for the studyThe authors. FundingThis study is partially supported by Merck KgA, Darmstadt, Germany. DisclosureE. Martinelli: Advisory / Consultancy: Amgen, Bayer, Merck, Roche, Sanofi, Servier. T. Troiani: Advisory / Consultancy: Amgen, Bayer, Merck, Novartis, Roche. F. Ciardiello: Advisory / Consultancy: Merck Kga, Bayer, Amgen, Rocher, Servier, Pfizer. All other authors have declared no conflicts of interest.

Antitumor effects of flavopiridol, a cyclin-dependent kinase inhibitor, on human cholangiocarcinoma in vitro and in an in vivo xenograft model

Flavopiridol, a pan-cyclin-dependent kinase (CDK) inhibitor, was recently identified as an effective antitumor agent for several cancers. We investigated the antitumor effect of flavopiridol on cholangiocarcinoma (CCA), in vitro and in vivo. A methylthiotetrazole assay revealed that the proliferation of certain CCA cells was inhibited by flavopiridol, which induced the caspase-dependent apoptosis of CCA cells. Although increased cell cycle arrest was observed at the G2/M phase, caspase activation occurred earlier than 24 h, indicating that caspase-dependent apoptosis is the major pathway for the suppression of cell proliferation. Flavopiridol potently reduced the CCA tumor growth in a xenograft model without observable adverse effects. These findings indicated that flavopiridol could be a potential antitumor agent for the treatment of CCA.

Phase II study of avelumab in combination with cetuximab in pre-treated RAS wild-type metastatic colorectal cancer patients: CAVE (cetuximab-avelumab) Colon.

TPS731 Background: The immune system plays a crucial role in modulating response to monoclonal antibodies therapy in cancer. Novel immunecheckpoint inhibitors have demonstrated potent efficacy alone and in combinations with cytotoxic agents in several cancers. In this regard, avelumab in combination with cetuximab might be a relevant rechallenge strategy in RAS wild-type (WT) metastatic colorectal cancer (mCRC) patients treated in first-line with chemotherapy (CT) in combination with anti-EGFR drugs and who achieved a complete or partial response. Methods: CAVE Colon is a single arm, multi-center phase II study designed to evaluate the efficacy of avelumab and cetuximab in pre-treated RAS WT mCRC patients. Eligible patients: pathologically confirmed RAS WT mCRC treated with a first-line CT in combination with an anti-EGFR agent with a major response achieved (complete or partial), who have progressed to a second line therapy, and received no prior immunotherapy. Primary endpoint is overall survival, secondary endpoint are overall response rate according to RECIST 1.1, progression free survival and safety profile. The current study seeks to demonstrate a median OS of 11 months (alternative hypothesis) by the experimental combination for comparison with historical median OS 8.0 (null hypothesis) with standard third line treatments, which correspond to an improvement of OS at six months from 40% to 57%. It was estimated that it would be needed to enroll 66 patients to achieve a 80% power with a one-sided 5% level test. The accrual period will be 18 months and the total duration of the study will be 36 months. Considering a potential drop-out of approximately 15% of patients, a total of 75 patients will be recruited. Seven patients have been enrolled and started treatment to date (September 15, 2018) with avelumab 10 mg/kg q14 as a one-hour i.v. infusion and cetuximab at 400 mg/m2 over two-hour and subsequently 250 mg/m2 q 14 as one-hour i.v. infusion until disease progression or unacceptable toxicity. EudraCT number: 2017-004392-32. This study is partially supported by Merck KgA, Darmstadt, Germany. Results: N/A. Conclusions: N/A. Clinical trial information: EudraCT number: 2017-004392-32.

4-Methylthio-3-butenyl isothiocyanate mediates nuclear factor (erythroid-derived 2)-like 2 activation by regulating reactive oxygen species production in human esophageal epithelial cells

4-Methylthio-3-butenyl isothiocyanate (MTBITC) extracted from daikon (Raphanus sativus), which shows antimutagenicity, may have applications as an effective chemopreventive agent in several cancers; however, few reports have described the associated mechanisms. We investigated whether MTBITC induced cytoprotective genes, including phase II enzymes, in Het-1A human esophageal epithelial cells. HMOX1, NQO1, and GCLC mRNA levels and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) protein levels were increased in Het-1A cells treated with 10 μM MTBITC. Reactive oxygen species (ROS) tended to increase when Het-1A cells were treated with MTBITC, and the increases in ROS and Nrf2 expression in the cells treated with MTBITC were completely abolished by treatment with N-acetyl-l-cysteine. We also examined the relationships between Nrf2 activation and mitogen-activated protein kinase (MAPK) signaling by western blot analysis. MTBITC induced extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 phosphorylation in Het-1A cells; however, MTBITC did not affect the relationship between Nrf2 activation and MAPK responses. In the present study, we found that MTBITC induced Nrf2 activation and cytoprotective genes via ROS production in Het-1A cells. These results suggest that MTBITC may have the potential for preventing esophageal carcinogenesis through modification of carcinogen metabolism by phase II enzyme induction via ROS production.

Targeting CD47 and Autophagy Elicited Enhanced Antitumor Effects in Non-Small Cell Lung Cancer.

CD47-specific antibodies and fusion proteins that block CD47-SIRPα signaling are employed as antitumor agents for several cancers. Here, we investigated the synergistic antitumor effect of simultaneously targeting CD47 and autophagy in non-small cell lung cancer (NSCLC). SIRPαD1-Fc, a novel CD47-targeting fusion protein, was generated and was found to increase the phagocytic and cytotoxic activities of macrophages against NSCLC cells. During this process, autophagy was markedly triggered, which was characterized by the three main stages of autophagic flux, including formation and accumulation of autophagosomes, fusion of autophagosomes with lysosomes, and degradation of autophagosomes in lysosomes. Meanwhile, reactive oxygen species and inactivation of mTOR were shown to be involved in autophagy initiation in SIRPαD1-Fc-treated cells, indicating a probable mechanism for autophagy activation after targeting CD47 by SIRPαD1-Fc. Inhibition of autophagy enhanced macrophage-mediated phagocytosis and cytotoxicity against SIRPαD1-Fc-treated NSCLC cells. In addition, simultaneously targeting both CD47 and autophagy in NSCLC xenograft models elicited enhanced antitumor effects, with recruitment of macrophages, activated caspase-3, and overproduction of ROS at the tumor site. Our data elucidated the cytoprotective role of autophagy in CD47-targeted therapy and highlighted the potential approach for NSCLC treatment by simultaneously targeting CD47 and autophagy. Cancer Immunol Res; 5(5); 363-75. ©2017 AACRSee related Spotlight by Kaufman, p. 355.

Abstract P4-21-18: Pre-clinical antitumor activity, tumor localization, and pharmacokinetics of MP0274, an apoptosis inducing, biparatopic HER2-targeting DARPin®

Abstract Background: HER2 positivity is an important predictive factor for treatment with anti-HER2 agents in several cancers. However, currently available monoclonal antibody and tyrosine kinase inhibitor drugs rarely achieve full disease control. We have developed a new HER2-targeting molecule with a unique pro-apoptotic mode of action that may provide additional benefit to patients. The DARPin® MP0274* binds to two distinct non-overlapping HER2 epitopes and to human serum albumin for half-life extension. As previously shown**, in vitro, MP0274 induces apoptosis and inhibits proliferation of cells expressing HER2 (IHC3+, IHC2+ and IHC1+) and potently inhibits HER2/HER3 downstream signaling. To support clinical development of MP0274, we tested the potency of MP0274 in several HER2 expressing patient-derived xenograft (PDX) models and investigated tumor localization. In addition, pharmacokinetics (PK) analysis was performed in cynomolgus monkeys. Methods: Antitumor activity of MP0274 was tested in breast and gastric HER2 expressing PDX mouse models and was compared to standard of care therapies. Tumor localization of MP0274 was studied using an Indium-111 labeled version of MP0274 in a human ovarian adenocarcinoma (SKOV-3) xenograft model by whole-body SPECT/CT imaging. The PK of MP0274 was studied in cynomolgus monkeys (MP0274 is cross-reactive with cynomolgus HER2). Results: In breast and gastric cancer PDX models, MP0274 showed superior efficacy compared to trastuzumab and lapatinib and equivalent efficacy compared to trastuzumab plus pertuzumab as measured by relative tumor volume. The imaging study with SPECT/CT demonstrated that MP0274 localizes effectively to the HER2-expressing human tumor within 24 h. The PK study in cynomolgus monkeys showed a half-life of ≥5 days at doses of 5 and 10 mg/kg while at the lowest dose tested (1 mg/kg) MP0274 had a terminal half-life of 0.4 days. The PK results are indicative of target-mediated clearance that becomes saturated at doses above 1 mg/kg. Conclusions: MP0274, with its unique pro-apoptotic mode of action, demonstrates excellent activity in preclinical PDX models, fast localization to tumor and a long half-life in cynomolgus monkeys. MP0274 was well tolerated in all studies. These results suggest that MP0274 has the potential to provide additional clinical benefit to patients with HER2-expressing tumors. A GLP repeated dose toxicology study is ongoing and a phase I clinical trial is in preparation. * DARPins are small repeat proteins, designed to bind targets with high affinity and specificity, which can be combined in a modular fashion to produce multi-functional agents. ** U. Fiedler et al. SABC 2013. Abstract# 1094 &amp; Poster# P4-12-30. Citation Format: Fiedler U, Metz C, Zitt C, Bessey R, Béhé M, Blanc A, Schibli R, Dolado I, Herbst J, Dawson KM, Kiemle-Kallee J. Pre-clinical antitumor activity, tumor localization, and pharmacokinetics of MP0274, an apoptosis inducing, biparatopic HER2-targeting DARPin® [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P4-21-18.

The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models.

Avoidance of apoptosis is critical for the development and sustained growth of tumours. The pro-survival protein myeloid cell leukemia 1 (MCL1) is overexpressed in many cancers, but the development of small molecules targeting this protein that are amenable for clinical testing has been challenging. Here we describe S63845, a small molecule that specifically binds with high affinity to the BH3-binding groove of MCL1. Our mechanistic studies demonstrate that S63845 potently kills MCL1-dependent cancer cells, including multiple myeloma, leukaemia and lymphoma cells, by activating the BAX/BAK-dependent mitochondrial apoptotic pathway. In vivo, S63845 shows potent anti-tumour activity with an acceptable safety margin as a single agent in several cancers. Moreover, MCL1 inhibition, either alone or in combination with other anti-cancer drugs, proved effective against several solid cancer-derived cell lines. These results point towards MCL1 as a target for the treatment of a wide range of tumours.