bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3869: A nanofiber-based biosensor for ultrasensitive detection of serum CRP in melanoma
Authors: Lin Zuan-Tao , Li Yaxi , Wu Tianfu
Published: August 2018

Purpose: Early detection of melanoma is important to improve survival; however, the technology for accurate early diagnosis is still challenging. This study is to develop a novel polymeric biosensor that could provide superior sensitivity in the detection of potential early biomarkers (e.g., CRP) of melanoma.

Experimental Procedure: Our strategy is to construct a highly specific molecular recognition core and a highly responsive transducer made of conductive polypyrrole (PPy) nanofibers. Briefly, a polymeric matrix was produced by polymerization of acrylamide (AM), methylenebisacrylamide (MBAA), N-Isopropylacrylamide (NIPAAm) and CRP/CRP-aptamer complex first. Next, taking advantage of the porous structure of this NIPAAm-AM-CRP-aptamer/CRP polymeric matrix, we synthesized a polypyrrole-based conductive nanofiber structure using copper phthalocyanine-3,4',4'',4‴-tetrasulfonic acid tetrasodium salt (CuPT) as a dopant counterion in situ to achieve uniformly dispersed nanofibers within the polymeric network of NIPAAm-AM-CRP-aptamer/CRP. After removing CRP recombinant protein, we were able to obtain a robust CuPT-PPy/NIPAAm-AM polymeric sensor for the detection of CRP with high sensitivity and selectivity. Two-step signal amplification cascades were involved in this CRP-specific polymeric sensor: 1) CRP binding-induced polymeric network shrinkage; 2) shrinkage-triggered conductance change of the polymeric network. Therefore, serum CRP levels could be quantitatively analyzed through monitoring the conductance change caused by polymeric network shrinkage upon aptamer-CRP binding.

Results: The limit of detection (LOD) of the polymeric sensor for detection of human recombinant CRP could reach 10^−19 M. The Fourier transform infrared (FT-IR) spectra confirmed the chemical structure of the polymeric network, and the morphology was observed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The results of FT-IR reveal that CuPT-PPy nanofiber was successfully synthesized in situ. The nanostructure of the nanofibers was clearly observed using SEM and AFM, indicating that the diameter of nanofiber is about 20 nm. This CRP-specific biosensor and a commercial CRP ELISA kit were used to perform side-by-side measurement of serum CRP in melanoma patients.

Conclusion: Our results indicated that this CRP-specific conductive polymeric senor is highly sensitive and selective in accurately discriminating melanoma patients from healthy controls. Such a flexible conductive polymeric biosensor may hold great promise as a point-of-care device in the diagnostics of melanoma or other cancers.

Citation Format: Zuan-Tao Lin, Yaxi Li, Tianfu Wu. A nanofiber-based biosensor for ultrasensitive detection of serum CRP in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3869.
Abstract 3878: Drug-loaded microparticles as a treatment approach for pancreatic cancer
Authors: Munoz-Sagastibelza Maria , Okome Oluwafeyikemi , Sanchez Jenny E. Paredes , Palileo Albert , Burkhart Catherine , Martello-Rooney Laura
Published: August 2018

Pancreatic cancer is the fourth leading cause of cancer death in the United States with only 7% of diagnosed patients surviving 5 years. Most pancreatic cancer patients are not surgical candidates due to advanced stage at diagnosis. Also, current systemic chemotherapies have not been very effective at decreasing tumor burden. Poly(lactic-co-glycolic acid)-based (PLGA) microparticles (MPs) are a promising tool for localized drug delivery within the tumor due to their biocompatibility, flexibility in drug encapsulation and extended drug release. The present study investigated whether gemcitabine-loaded microparticles (GMPs), paclitaxel-loaded microparticles (PMPs) or sequential treatment of both, in comparison with blank (no drug) MPs, systemic treatments and no treatment control, are able to promote cancer cell killing effects and modulate drug resistance in vitro and in vivo. We previously showed in vitro studies with PANC-1 cells and we were able to complete the MPs studies with a second human pancreatic cancer cell line, MIAPaCa-2. In both cases, we tested the effect of single and combination treatments on two well-known resistance markers for gemcitabine, ribonucleotide reductase catalytic subunit M1 and cytidine deaminase, as well as on the promotion of cell death measuring cleaved caspase-3 (CC3). When treated with GMPs alone, both markers went up suggesting an increase in resistance against gemcitabine. Interestingly, both cell lines treated with PMPs alone showed an increase in CC3 expression and a significant decrease in the expression of both markers. Subsequently, we tested the in vivo efficacy of MPs by direct injection into established subcutaneous MIAPaCa-2 tumors in nude mice. Following four weeks of treatment, the tumors were excised, biopsied for protein analysis and frozen in OCT to allow visualization of fluorescent MPs and detect apoptosis by immunofluorescence. We also want to evaluate reactive oxygen species production; where we expect to see an increase in the MPs treated groups. This analysis is in progress. In conclusion, we observed a decrease in cell viability and drug resistance proteins in vitro using the drug-loaded microparticles in two grade 3 (or high grade) pancreatic cancer cell lines. The described drug delivery method has the potential to be a more efficient treatment modality than systemic gemcitabine and paclitaxel against early stage and locally advanced pancreatic cancer.

Citation Format: Maria Munoz-Sagastibelza, Oluwafeyikemi Okome, Jenny E. Paredes Sanchez, Albert Palileo, Catherine Burkhart, Laura Martello-Rooney. Drug-loaded microparticles as a treatment approach for pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3878.
Abstract 3879: Enhancement of tumor penetration by drug-loaded nanoparticles: An innovative targeted strategy for neuroblastoma
Authors: Ponzoni Mirco , Brignole Chiara , Emionite Laura , Bruno Silvia , Guarnieri Daniela , Sitia Leopoldo , Bauckneht Matteo , Buschiazzo Ambra , Rossi Andrea , Paolo Daniela Di , Perri Patrizia , Curnis Flavio , Gori Alessandro , Sementa Angela Rita , Cilli Michele , Pompa Pier Paolo , Sambuceti Gianmario , Corti Angelo , Pastorino Fabio
Published: August 2018

Background: Anti-cancer drugs-loaded targeted nanocarriers can reduce side-effects and improve therapeutic efficacy in preclinical studies. However, poorly perfused and dysfunctional tumour vessels limit the transport of the payload into the parenchyma of solid tumours. The use of tumour-penetrating nanocarriers might enhance tumour penetration and anti-tumour effects.

Methods: A consensus motif, mediator of tissue penetration (TP) was added to a previously characterized neuroblastoma (NB)-targeting peptide (pep). In vitro NB cell association and internalization of TP-pep, either free or coupled to Stealth Liposomes (SL), were tested by FACS and confocal microscopy. In vitro cytotoxic potential of a novel doxorubicin (DXR)-loaded liposomal (TP-pep-SL[DXR]) was evaluated by MTS assay. Three mouse xenograft models mimicking the growth and spread of NB in humans (injection routes: subcutaneous; adrenal gland; tail vein) were enrolled to examine in vivo penetration, vascular permeability, tumour glucose consumption and sensitivity in response to TP-pep-SL[DXR].

Results: Compared to pep, TP-pep increases its cellular association in vitro on cell lines and cells derived from NB patients. When coupled to SL, TP-pep enhances liposomes penetration and cytotoxic effects in vitro and increases binding and penetration in a mouse model of NB. Moreover, in vivo accumulation of Evans Blue dye within the tumour mass reveals that TP-pep-SL[DXR] increases the tumour vascular permeability into NB tumour mass, but not in non-tumour tissues. Compared to pep-targeted liposomes, TP-pep-SL[DXR] leads to an increased anti-NB effect towards all the animal models tested.

Conclusion: Our findings demonstrate that the enhancement of tumour penetration by drug-loaded nanoparticles might represent an innovative targeted strategy for NB.

Citation Format: Mirco Ponzoni, Chiara Brignole, Laura Emionite, Silvia Bruno, Daniela Guarnieri, Leopoldo Sitia, Matteo Bauckneht, Ambra Buschiazzo, Andrea Rossi, Daniela Di Paolo, Patrizia Perri, Flavio Curnis, Alessandro Gori, Angela Rita Sementa, Michele Cilli, Pier Paolo Pompa, Gianmario Sambuceti, Angelo Corti, Fabio Pastorino. Enhancement of tumor penetration by drug-loaded nanoparticles: An innovative targeted strategy for neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3879.
Abstract 3880: Peptide-targeted chemotherapy for pancreatic cancer
Authors: Lin Chin-Tarng , Wu Chen-Der , Lee Jen-Chieh , Wu Han-Chung , Lee Chung-Wei , Lin Chin-Feng , Hsu Ming-Chieh
Published: August 2018

The purpose of this abstract was to assess the efficacy of peptide targeted chemotherapy for pancreatic carcinoma. Previously we have constructed four peptides that bind specifically to cancer cell lines, which were derived from three different carcinoma cell lines and their vascular endothelia: L-peptide: (L-P, anti-cancer cell membrane), RLLDTNRPLLPY; SP-94-P (anti-hepato-pancreatic cancer cell membrane), SFSHHTPILP; PC5-52-peptide (anti-tumor endothelia), SVSVGMKPSPRP; and control peptide, RLLDTNRGGGGG. These peptides were linked to pegylated liposomal iron oxide nanoparticles to identify the targeted tumor cells and vascular endothelia using MRI analysis, and were also linked to dextran coated liposomal doxorubicin (L-D) for treatment of non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice bearing pancreatic cancer cell (PANC-1) xenografts. Our results demonstrated that the tumor intensity of MRI is clearly decreased after SP-94-P-iron oxide was applied, and that in combination of application of L-P linked L-D (L-P-L-D) plus SP-94-P linked L-D (SP-94-P-L-D) and PC5-52-P linked L-D (PC5-52-P-L-D), they could inhibit pancreatic tumor growth with very mild adverse events. The use of the control peptide linked L-D also led to a decrease of the xenograft size, but also induced marked apoptotic change of their visceral organs. It is concluded that the combination of L-P-L-D, SP-94-P-L-D and PC5-52-P-L-D to treat pancreatic cancer xenograft in NOD SCID mice can clearly inhibit pancreatic cancer growth with minimal adverse events.

Citation Format: Chin-Tarng Lin, Chen-Der Wu, Jen-Chieh Lee, Han-Chung Wu, Chung-Wei Lee, Chin-Feng Lin, Ming-Chieh Hsu. Peptide-targeted chemotherapy for pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3880.
Abstract 3882: Functional validation of a genome-wide association study (GWAS) SNP associated with anthracycline-induced congestive heart failure
Authors: Wu Xi , Xue Gloria , Gardner Laura , Shen Fei , Jiang Guanglong , Philips Santosh , Cunningham Geneva , Schneider Bryan P.
Published: August 2018

Anthracyclines are commonly used chemotherapies for the treatment of breast cancer, but they can cause dose-related cardiotoxicities and lead to congestive heart failure (CHF) in ~2% of patients. There are no clinically available predictive biomarkers for this toxicity. Previously we have identified and validated the association of a single nucleotide polymorphism (SNP), rs28714259, with CHF in three large adjuvant phase III breast cancer trials. rs28714259 locates in a putative glucocorticoid receptor (GR) binding site and the risk (minor) allele is predicted to disrupt GR binding. Interestingly, activation of GR-signaling by dexamethasone has been shown to protect cardiomyocytes from doxorubicin-induced apoptosis in rat. To test whether the risk allele affects GR binding efficiency in vitro, we performed electrophoretic mobility shift assay (EMSA) in MCF-7 and A549 cells. 100nM of dexamethasone for 4 hours induced a prominent band shift in either the major- or risk-allele probes. Notably, we observed a ~50% reduction in the shifted band intensity of risk-allele probes compared to major-allele probes, suggesting decreased binding in risk-allele probes. No additional supershift was detected after the binding reaction was co-incubated with a GR antibody. However, the previously shifted band was significantly diminished in both the major- and risk-allele probes, probably due to a conformational change induced by GR antibody and leading to the disruption of GR/DNA binding, which in turn suggests the observed shift was specifically caused by GR. Further validations are ongoing using cardiomyocytes derived from human induced pluripotent stem cells (iPSCs). These studies provide further insight into the function and role of rs28714259 in anthracycline-induced CHF.

Citation Format: Xi Wu, Gloria Xue, Laura Gardner, Fei Shen, Guanglong Jiang, Santosh Philips, Geneva Cunningham, Bryan P. Schneider. Functional validation of a genome-wide association study (GWAS) SNP associated with anthracycline-induced congestive heart failure [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3882.
Abstract 3893: Quantitative contribution of rs75017182 to dihydropyrimidine dehydrogenase mRNA splicing and enzyme activity
Authors: Nie Qian , Shrestha Shikshya , Tapper Erin E. , Trogstad-Isaacson Colbren S. , Bouchonville Kelly J. , Offer Steven M. , Diasio Robert B.
Published: August 2018

5-Fluorouracil (5-FU) has remained a widely prescribed anti-cancer drug for decades. However, 5-FU–based treatments show inter-individual variability, with up to 34% of 5-FU–treated patients developing severe (grade 3+) adverse toxicity. Dihydropyrimidine dehydrogenase (DPD; DPYD gene) is the rate-limiting enzyme of 5-FU catabolism and converts 80–85% of 5-FU into its inactive metabolites. Three deleterious genetic variations in DPYD are well-established predictors of severe 5-FU toxicity (i.e., *2A, p. I560S, and p. D949V); however, these variants only explain 30–35% of adverse cases. Recently, an intronic variant (rs75017182) has been suggested to contribute to 5-FU–related toxicity by promoting the alternative splicing of DPYD. However, clinical studies are conflicting as to the degree to which rs75017182 affects DPD enzyme function, and the true contribution of the rs75017182 variant towards DPD function and 5-FU toxicity remains unclear and inconclusive. In the present study, we directly examined the effect of the intronic SNP rs75017182 on DPYD mRNA splicing, DPYD expression, and DPD enzyme function. We genotyped 3950 healthy Caucasian volunteers from the Mayo Clinic Biobank to identify the rs75017182, *2A, p.I560S, and p.D949V variants. From the initial cohort, 204 volunteers who were rs75017182 heterozygous carriers were recruited as our study population. Alternative splicing and DPD activity in rs75017182 carriers were measured. The findings were confirmed using a novel mini-gene reporter system in vitro. A moderate, but significant, reduction (30%) was detected in canonically spliced DPYD expression in rs75017182 carriers compared to non-carriers. A correlative reduction (35%) in DPD enzyme activity was observed in those carriers, which was similar to that of D949V carriers (31%). The results demonstrated an association of this deep intronic variant with decreased DPD activity and suggested that rs75017182 may be a predictor of 5-FU toxicity similar to D949V.

Citation Format: Qian Nie, Shikshya Shrestha, Erin E. Tapper, Colbren S. Trogstad-Isaacson, Kelly J. Bouchonville, Steven M. Offer, Robert B. Diasio. Quantitative contribution of rs75017182 to dihydropyrimidine dehydrogenase mRNA splicing and enzyme activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3893.
Abstract 3901: Characterizing antibody-drug conjugate cytotoxicity using four different real-time assays
Authors: Niles Andrew L. , Kupcho Kevin , Duellman Sarah , Vidugiriene Jolanta , Lazar Dan , Cali James J.
Published: August 2018

Recent clinical successes using antibody drug conjugates (ADC) directed to hematological or solid tumor cancers have re-invigorated immunotherapeutic drug development interest. Unfortunately, efficacious ADC are inherently complex entities comprised of a targeting antibody, a stable but cleavable linker, and a toxic payload, for which the best combinations are often identified through iterative trial and error. Therefore, an efficient and systematic method for rank ordering ADC candidates is sought to evaluate their intrinsic pharmacodynamic properties. We chose SKBR3 (Her2+) and T47D (Her2-) cell models to define the on- and off-target cytotoxic potential of trastuzumab emtansine. We employed a suite of four, newly developed real-time assays to measure exposure-dependent changes in viability, caspase activation, phosphatidylserine (PS) exposure and loss of membrane integrity to define potency and magnitude of response, as well as primary mode of cell death. We discovered that trastuzumab emtansine mediated dose-dependent effects on SKBR3, with cytostasis beginning at 17h near 1μg/ml, caspase induction at 20h, which led to PS exposure at about 21h and secondary necrosis at 30h. During this same time course, full dose-response curves were generated for each biomarker as EC50 values evolved and matured to their respective maximal responses. Conversely, the T47D non-target cell population exhibited only minor cytotoxicity at the latest time points, presumably owing to linker degradation and non-specific payload release. Taken together, these data define the in vitro pharmacological disposition of trastuzumab emtansine, including the kinetic component of functionality related to antigen/antibody engagement, internalization, linker mediated payload release and resulting toxicity. This study may provide a tangible roadmap for the critical evaluation of other ADC under development.

Citation Format: Andrew L. Niles, Kevin Kupcho, Sarah Duellman, Jolanta Vidugiriene, Dan Lazar, James J. Cali. Characterizing antibody-drug conjugate cytotoxicity using four different real-time assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3901.
Abstract 3902: A novel biologic ADI-TRAIL fusion protein benefits from structural and functional complementarity of its components arginine deiminase and TRAIL, induces cancer cell apoptosis in vitro, and inhibits tumor growth in vivo
Authors: Brin Elena , Wu Katherine , He Yudou , Shia Wei-Jong , Kuo Mario M. , Chen Li-Chang , Dagostino Eleanor , Hickey Richard , Almassy Bob , Showalter Richard , Thomson Jim
Published: August 2018

TNF-related apoptosis-inducing ligand (TRAIL) receptor agonists have shown good safety profile and efficacy in preclinical cancer models. However, clinical success has been limited due to poor PK and/or development of resistance to death receptor-induced apoptosis. To address these issues we have conceived and produced a fusion protein of arginine deiminase (ADI) and TRAIL. ADI is an enzyme that converts arginine into citrulline. Arginine deprivation can inhibit growth of arginine auxotrophic cancers that lack key enzymes enabling normal cells to produce arginine from citrulline. ADI is currently being evaluated in clinical trials and is well-tolerated. We (and others) observed synergy between ADI and TRAIL in inducing apoptosis in a number of cancer cell lines including those that are otherwise resistant to rhTRAIL. We have explored potential mechanisms of synergy and will present our findings. Using in silico modeling we predicted structural complementarity between ADI and TRAIL with each modality stabilized when part of the fusion protein. We were able to produce multiple ADI-TRAIL fusion protein variants and have tested their activity in enzymatic and cell-based assays. Our experimental data confirmed in silico predictions of the structural advantages for ADI and TRAIL in the fusion protein. ADI enzymatic activity improved when part of the fusion protein and the fusion protein appeared to have higher potency than the combination of the two separate proteins in inducing cancer cell death. In mice ADI-TRAIL fusion protein exhibited extended half-life and was efficacious in HCT116 xenograft model, superior to rhTRAIL administered at the same molar amounts. Serum concentrations of the fusion protein had an inverse correlation with the tumor volume. ADI-TRAIL is a novel biologic that benefits from structural and functional synergies between its components and is showing promise as a cancer therapeutic in preclinical studies.

Citation Format: Elena Brin, Katherine Wu, Yudou He, Wei-Jong Shia, Mario M. Kuo, Li-Chang Chen, Eleanor Dagostino, Richard Hickey, Bob Almassy, Richard Showalter, Jim Thomson. A novel biologic ADI-TRAIL fusion protein benefits from structural and functional complementarity of its components arginine deiminase and TRAIL, induces cancer cell apoptosis in vitro, and inhibits tumor growth in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3902.
Abstract 3926: Discovering a novel microtubule inhibitor overcoming tumor multidrug resistance
Authors: Ning Nannan , Wu Chengyong , Wu Min , Yu Yamei , Zhu Changjun , Benes Cyril , Zhang Jianming , Deng Xianming , Chen Qiang , Ren Ruibao
Published: August 2018

Microtubule inhibitors, as chemotherapeutic drugs, are widely used for cancer treatment in clinic. However, the development of multidrug resistance (MDR) in cancer is a major challenge for microtubule inhibitors in its clinical implementation. From a high throughput drug screening using cells transformed by oncogenic RAS, we identified a small molecule AR8-1006 that blocks cell proliferation. The analysis of structure-activity relationship indicated that this serial of scaffold represents a potent inhibitor of tumor cell growth. AR8-1006 shows activities against a larger panel of more than 1000 human cancer cell lines with a wide variety of tissue origins. Further analysis showed that this compound depolymerizes microtubules and affects formation of the spindle. Interestingly, it induces the spike-like conformation of microtubules in vitro and in vivo, which is different from typical microtubule modulators. Structural analysis revealed that this serial of compounds binds to the colchicine pocket at the intra-dimer interface, though mostly not overlapping with colchicine binding. More importantly, AR8-1006 displays favorable pharmacological properties on overcoming tumor MDR both in vitro and in vivo. Taken together, our data reveal a novel scaffold represented by AR8-1006 that could be developed as cancer therapeutics, especially for tumor multidrug resistance.

Citation Format: Nannan Ning, Chengyong Wu, Min Wu, Yamei Yu, Changjun Zhu, Cyril Benes, Jianming Zhang, Xianming Deng, Qiang Chen, Ruibao Ren. Discovering a novel microtubule inhibitor overcoming tumor multidrug resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3926.
Abstract 3932: Metabolic biomarkers predict the sensitivity to phosphatidylinositol 3-kinase pathway inhibitor in small-cell lung carcinoma
Authors: Umemura Shigeki , Makinoshima Hideki , Ishii Genichiro , Sasaki Takehiko , Esumi Hiroyasu , Goto Koichi , Tsuchihara Katsuya
Published: August 2018

Comprehensive genomic analysis has revealed that the PI3K/AKT/mTOR pathway is a feasible therapeutic target in small-cell lung carcinoma (SCLC). However, biomarkers to identify patients likely to benefit from inhibitors of this pathway have not been identified. Here we show that metabolic features determine sensitivity to the PI3K/mTOR dual inhibitor gedatolisib in SCLC cells. We analyzed the phosphatidyl lipid profile including the acyl groups of 11 SCLC cell lines using electrospray ionization mass spectrometry. We found that a specific phosphatidylinositol (3,4,5)-triphosphate (PIP3) subspecies lipid product, PIP3 (38:4), is predictive in assessing sensitivity to gedatolisib. We generated a large-scale metabolomic dataset for 28 SCLC cell lines. Notably, we found that higher amounts of purine-related aqueous metabolites such as hypoxanthine led to resistance to PI3K pathway inhibition. The levels of the mRNA encoding hypoxanthine phosphoribosyl transferase 1 (HPRT1), a key component of the purine salvage pathway, differed significantly between SCLC cells sensitive or resistant to gedatolisib. Loss of HPRT1 in SCLC cells decreased cellular proliferation in the presence of gedatolisib compared with a control. Further, complementation with purine metabolites could reverse the vulnerability to targeting of the PI3K pathway in SCLC cells normally sensitive to gedatolisib. These results indicate that the mechanism of resistance to PI3K pathway inhibitors is mediated by activation of the purine salvage pathway, which supplies purine resources for nucleotide biosynthesis. Metabolic profiles of clinical samples of SCLC revealed that purine-related metabolites were higher in SCLC tumor tissues, suggesting that higher amounts of purine-related aqueous metabolites might be characteristic of SCLC biology. Metabolomics is a powerful approach for identifying novel therapeutic biomarkers applicable to SCLC treatment.

Citation Format: Shigeki Umemura, Hideki Makinoshima, Genichiro Ishii, Takehiko Sasaki, Hiroyasu Esumi, Koichi Goto, Katsuya Tsuchihara. Metabolic biomarkers predict the sensitivity to phosphatidylinositol 3-kinase pathway inhibitor in small-cell lung carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3932.
Abstract 3933: Circadian timing regimen for alpelisib (NVP-BYL719), a selective inhibitor of the class Ia PI3K isoform alpha to maximize therapeutic index
Authors: Schnell Christian R. , Ferrat Thomas , Wyss Daniel , Tinetto Walter , Tobler Sonja , Fritsch Christine , Jensen Michael
Published: August 2018

The gene encoding for the catalytic subunit of the phosphatidylinositol-3-kinase (PI3K), p110a (PIK3CA) is the most frequently mutated kinase in cancer. This discovery triggered the development of small molecule anti-PI3K inhibitors, such as NVP-BYL719. However, the PI3K / Akt signaling pathway plays not only an important role in promoting cell growth, proliferation and survival but also in regulating glucose homeostasis by directly mediating insulin-stimulated glucose uptake into insulin sensitive tissues (adipocytes and muscles). NVP-BYL719 (a selective inhibitor of the class Ia PI3K isoform alpha) given in the morning is facing on-target tolerability challenge (hyperglycemia) in clinical trials limiting the dose being administered to patient (1). As glucose metabolism is highly impacted by circadian rhythms in patients and rodents, we have decided to adopt an integrative circadian-timing approach in our pre-clinical models to interrogate the benefit of morning vs evening dosing for BYL719. By using a newly developed radio-telemetry technology (2), we were able to record in real time and 'around-the-clock' blood glucose levels in stress-free, freely moving rats dosed with NVP-BYL719 at different regimens. The dynamic profile of hyperglycemia observed after active or inactive period dosing of NVP-BYL719 (50 mg/kg qd p.o.) were similar. Dosing before the inactive phase (10 a.m.) allowed blood glucose to normalize in between 2 doses, which could not be achieved when dosing before the active phase (5 p.m.). After treatment discontinuation a significant hyperglycemia remained for a period up to 12h in the group dosed before the active phase (5 p.m.). Circadian “evening” NVP-BYL719 dosing is associated with a better control of glycaemia. Clinically this could potentially translate to better compliance and longer time on treatment hence better efficacy for patient. Based on these findings we could recommend optimized treatment schedules for future combination experiments in the clinic with NVP-BYL719. 1) Juric et al, “Phase I study of the PI3Kα Inhibitor BYL719, as a Single Agent in Patients with Advanced Solid Tumors (AST)”, Annals of Oncology (2014), 25 (Supp. 4) 2) Brockway et al, “Fully Implantable Arterial Blood Glucose Device for Metabolic Research Applications in Rats for Two Months”, J Diabetes Sci Technol (2015), 9(4):771-81

Citation Format: Christian R. Schnell, Thomas Ferrat, Daniel Wyss, Walter Tinetto, Sonja Tobler, Christine Fritsch, Michael Jensen. Circadian timing regimen for alpelisib (NVP-BYL719), a selective inhibitor of the class Ia PI3K isoform alpha to maximize therapeutic index [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3933.
Abstract 3936: Bromodomain testis-specific protein BRDT is expressed in a subset of esophageal squamous cell carcinomas and controls expression of differentiation-associated genes
Authors: Wang Xin , Hamdan Feda H. , Sen Madhobi , Kutschat Ana P. , Johnsen Steven A.
Published: August 2018

Esophageal cancer is one of the most malignant cancers, ranking as the sixth leading cause of cancer-related deaths worldwide. The poor survival rate and prognosis highlight the limitations in the biological understanding of esophageal cancer and the urgent need for identification of novel targeted molecular therapies. Recently, large scale genomic analyses have revealed the extensive alternations of epigenetic regulators which may be used as a basis for developing new “epigenetic drugs “. BRDT, bromodomain testis-specific protein, is a member of the bromodomain and extra-terminal (BET) family of epigenetic reader proteins. BET proteins can regulate gene expression by recognizing acetylated lysines, thus playing important roles in both normal development and disease progression. Inhibition of BET proteins has emerged as a potential therapy for many types of cancer. In normal human tissues, unlike the other members of BET family, BRDT is exclusively highly expressed in testes where it drives the meiotic and post-meiotic gene expression to promote spermatogenesis. We have identified BRDT to be expressed in over 20% of esophageal squamous cell carcinoma (ESCC), a predominant subtype of esophageal cancer. Knockdown of BRDT does not affect cell proliferation, but leads to alterations in the expression of differentiation markers. In addition, RNA-seq following BRDT knockdown also supports a role of BRDT in cell differentiation. Depletion of BRDT does not alter the cellular response to BET inhibition. Surprisingly, we also identified BRDT transcripts encoding two truncated isoforms lacking the first bromodomain, which could potentially alter its epigenetic reader function. The genome occupancy profile of BRDT showed a significant enrichment at transcription start site (TSS), which is distinct from BRD4. It also showed potential interactions between BRDT and E2F family and SMAD family. Together, we showed that BRDT is overexpressed in ESCC cell lines and may influence ESCC development or progression in a subset of tumors by regulating cell differentiation-associated genes.

Citation Format: Xin Wang, Feda H. Hamdan, Madhobi Sen, Ana P. Kutschat, Steven A. Johnsen. Bromodomain testis-specific protein BRDT is expressed in a subset of esophageal squamous cell carcinomas and controls expression of differentiation-associated genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3936.
Abstract 3956: Omomyc-based cell-penetrating peptides: From proof of concept to a clinically viable anti-Myc therapy
Authors: Beaulieu Marie-Eve , Jauset Toni , Massó-Vallés Daniel , Martínez-Martín Sandra , Rahl Peter , Maltais Löika , Zacarias-Fluck Mariano F. , Casacuberta-Serra Sílvia , Pozo Erika Serrano del , Fiore Christopher , Foradada Laia , Cano Virginia Castillo , Guenther Matthew , Sanza Eduardo Romero , Oteo Marta , Tremblay Cynthia Tremblay , Martín Génesis , Letourneau Danny , Montagne Martin , Alonso Miguel Ángel Morcillo , Whitfield Jonathan R. , Lavigne Pierre , Soucek Laura
Published: August 2018

Deregulation of the MYC oncoprotein drives tumorigenesis in most–if not all–cancers and generally correlates with poor prognosis, suggesting that its inhibition would be a useful therapeutic strategy. Indeed, we have shown that Myc inhibition displays extraordinary therapeutic benefit in various transgenic mouse models of cancer (i.e., skin, lung, pancreatic cancer and glioma), without eliciting resistance to therapy, and causes only mild, well-tolerated and reversible side effects in normal tissues. For these studies we employed a dominant negative inhibitor of Myc, called Omomyc, which is an effective inhibitor of Myc transactivation function both in vitro and in vivo. Omomyc has so far been utilized exclusively as a transgene, as a successful proof of principle whose application was believed to be solely limited to gene therapy. Here, though, we show that the purified Omomyc polypeptide itself spontaneously transduces into cancer cells and effectively interferes with MYC transcription, abrogating cell cycle and promoting apoptosis in different cancer cells, independently of their mutational profile. Efficacy of the Omomyc polypeptide in two complementary murine models of non-small cell lung cancer (NSCLC) establishes its therapeutic potential through both direct tissue delivery (intranasal) and systemic intravenous administration, providing for the first time evidence that the Omomyc polypeptide is an effective MYC inhibitor worthy of clinical development.

Citation Format: Marie-Eve Beaulieu, Toni Jauset, Daniel Massó-Vallés, Sandra Martínez-Martín, Peter Rahl, Löika Maltais, Mariano F. Zacarias-Fluck, Sílvia Casacuberta-Serra, Erika Serrano del Pozo, Christopher Fiore, Laia Foradada, Virginia Castillo Cano, Matthew Guenther, Eduardo Romero Sanza, Marta Oteo, Cynthia Tremblay Tremblay, Génesis Martín, Danny Letourneau, Martin Montagne, Miguel Ángel Morcillo Alonso, Jonathan R. Whitfield, Pierre Lavigne, Laura Soucek. Omomyc-based cell-penetrating peptides: From proof of concept to a clinically viable anti-Myc therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3956.
Abstract 3984: Combination S6K and TAM tyrosine kinase targeting in PTEN-deficient glioma
Authors: Ennis Kelli N. , Sarma Pranjal , Behrmann Catherine A. , Wang Jie , Plas David R.
Published: August 2018

Loss of PTEN is one of the most frequent driver events in primary glioblastoma, conferring the ability to reconfigure metabolism and enhance apoptosis resistance to transformed glial cells. The ribosomal S6 protein kinases (S6Ks) are hyperactivated in PTEN-deficient settings, and genetic inactivation studies have shown that S6K1 deficiency can reduce the oncogenic growth of PTEN-deficient neoplasia. Investigating pharmacologic agents, we found that the S6K inhibitor LY-2584702 selectively elicits cytotoxic responses in PTEN-deficient cells when combined with the TAM kinase inactivating agent BMS-777607. Combined knockdown of TAM kinases sensitized cells to single agent treatment with LY-2584702. Conversely, expression of activated S6K1 or elevated AXL protected from combination treatment. These results indicate that both the TAMS and S6K1 are key regulators of survival in PTEN deficient cells.

Citation Format: Kelli N. Ennis, Pranjal Sarma, Catherine A. Behrmann, Jie Wang, David R. Plas. Combination S6K and TAM tyrosine kinase targeting in PTEN-deficient glioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3984.
Abstract 3989: Dual targeting of ErbB2/ErbB3 for treatment of SCL3A2-NRG1-mediated lung cancer
Authors: Jo Jeong Yeon , Shin Dong Hoon , Han Ji-Youn
Published: August 2018

We previously identified an oncogenic fusion gene involving the 5′ region of SLC3A2 and the 3′ region of NRG1 in a patient with invasive mucinous adenocarcinoma (IMA) of the lung. Here, we characterized the SLC3A2-NRG1 fusion gene in non-small cell lung cancer (NSCLC) and established an effective therapy for patients with SLC3A2-NRG1 fusion-positive cancer. The SLC3A2-NRG1 fusion product was composed of the SLC3A2 transmembrane domain and the epidermal growth factor (EGF)-like domain of the neuregulin 1 (NRG1) protein. The NRG1 family is classified as a ligand of the ERBB family. We identified ERBB3 and ERBB4 in the ERBB family as binding partners of the SLC3A2-NRG1 fusion protein via ligand and receptor binding assays. We confirmed that SLC3A2-NRG1 increased formation of a heterocomplex of ERBB3, which lacks tyrosine kinase activity, with ERBB2 using immunofluorescence staining, FACS analysis, and immunoprecipitation. Activation of the ERBB2-ERBB3 heterocomplex by SLC3A2-NRG1 increased colony formation and tumor growth in vivo in a xenograft model through the downstream signaling pathways of PI3K-AKT and MAP kinase. siRNA specific for ERBB2 and ERBB3, pertuzumab (a monoclonal antibody for ERBB2), lumretuzumab (a monoclonal antibody for ERBB3), and afatinib (an ERBB2 kinase inhibitor) all decreased ERBB2-ERBB3 heterocomplex formation, phosphorylation of each protein, and their downstream signaling. In addition, single treatment with pertuzumab, lumretuzumab, or apatinib decreased tumor volume and weight, whereas combination treatment with these drugs and paclitaxel enhanced generation of cleaved-caspase 3, PARP, and TUNEL-positive cells compared with each single treatment. Thus, the SLC3A2-NRG1 fusion gene plays an important role in lung cancer cell proliferation and tumor growth by promoting generation of the ERBB2-ERBB3 heterocomplex, its phosphorylation, and activation of the PI3K/ERK/mTOR signaling pathway. Inhibition of either ERBB2 or ERBB3 alone did not completely shut down downstream signaling of ERBB2 and ERBB3; however, inhibition of both ERBB2 and ERBB3 blocked downstream signaling activated by SLC3A2-NRG1 fusion. ERBB2 and ERBB3 might be promising targets for treatment of SLC3A2-NRG1-positive tumors.

Citation Format: Jeong Yeon Jo, Dong Hoon Shin, Ji-Youn Han. Dual targeting of ErbB2/ErbB3 for treatment of SCL3A2-NRG1-mediated lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3989.
Abstract 3992: Varlitinib demonstrates tumor regression and vessel normalization in ErbB-dependent and mutated beta-catenin hepatocellular carcinoma patient-derived xenograft model
Authors: Shuen Wai Ho , Ong Richard , Yeo Chloe , Banu Rebecca , Koh Lip Seng , Chee Chit Lai , Seet Qihui , Ooi Lisa , McHale Mark , Lindmark Bertil , Toh Han Chong , Hung Huynh
Published: August 2018

Hepatocellular carcinoma (HCC) is the most common form of liver cancer and current therapies such as sorafenib, regorafenib and lenvatinib, prolong median overall survival by less than a year. Hence, there is an urgent need for additional HCC therapies. Varlitinib is a potent, reversible, small molecule pan – HER inhibitor that selectively binds to EGFR, HER2 and HER4. Varlitinib is currently being clinically tested across multiple indications including biliary tract cancer and gastric cancer. In HCC patient – derived xenograft (PDX) models, varlitinib has been shown to mediate potent anti – tumour activities (ASCO 2016). To understand the molecular profiles of HCC PDX sensitive to varlitinib, we studied the ErbB family signalling in 56 HCC PDX models and varlitinib was found to be efficacious in HCC PDX with activated ErbB2 / 3 as well as mutated beta – catenin. In HCC PDX models with activated ErbB2 / 3 , treatment with varlitinib inhibited multiple proliferation and anti – apoptosis pathways including AKT, PI3K and Survivin pathways. In addition, normalisation of blood vessels by varlitinib in the PDX models, as assessed by staining with biotinylated lectin and anti – CD31, also reduced hypoxia in the tumour microenvironment as well as enhanced the infiltration of immune cells. Activating mutations in the Wnt / beta – catenin pathway lead to tumour development in HCC and occur in a subset of HCC tumours. HCC PDX models with beta – catenin mutation were found to be sensitive to varlitinib. Varlitinib led to down – regulation of mutated beta – catenin signalling, and enhanced staining of beta – catenin at the membrane instead of in the nucleus. Since activating mutations in the Wnt / beta – catenin pathway are detected in 26 – 40 % of HCC samples, our data shows that varlitinib may represent an alternative treatment for a significant subset of HCC which are ErbB / beta – catenin – dependent.

Citation Format: Wai Ho Shuen, Richard Ong, Chloe Yeo, Rebecca Banu, Lip Seng Koh, Chit Lai Chee, Qihui Seet, Lisa Ooi, Mark McHale, Bertil Lindmark, Han Chong Toh, Huynh Hung. Varlitinib demonstrates tumor regression and vessel normalization in ErbB-dependent and mutated beta-catenin hepatocellular carcinoma patient-derived xenograft model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3992.
Abstract 3994: Anti-metastatic drug MBQ-167 regulates cell polarity via Cdc42
Authors: Rivera-Robles Michael John , Medina-Velázquez Julia , Asencio-Torres Gabriela M. , González-Crespo Sahily , Rymond Brian C. , Rodríguez-Medina José , Dharmawardhane Suranganie
Published: August 2018

Cdc42 (cell division control protein 42 homolog) is a Rho GTPase highly conserved among eukaryotes in structure and function. Mechanical or chemical cues in the microenvironment stimulate the localized activation of Cdc42, aiding cells to polarize and arrange their components accordingly. Its role in cell polarity was first described in budding yeast and subsequently in other eukaryotes, including animal cells determining the direction of motility. Accordingly, in cancer Cdc42 mediates migration, invasion, and spread of tumor cells. Therefore, we target Cdc42 as therapeutic strategy to treat metastatic breast cancer. Our group designed and characterized the small molecule MBQ-167 as a potent inhibitor against Cdc42 activation (IC50=0.1 µM), demonstrating its effectiveness in preventing tumor growth and cell migration in vitro and in vivo. Because in yeast S. cereviciae Cdc42 is well known for regulating the location of budding events, we used this well-characterized model of cell polarization to test our previous findings of MBQ-167 and its specificity for Cdc42. By using a tetracycline-controlled transcriptional inactivation (Tet-Off) system at the promoter region of the CDC42 gene of yeast, we knocked down Cdc42 supplementing the media with doxycycline (DOX; IC=10ug/uL). Next, we analyzed the growth, budding pattern, and Cdc42 activity of budding yeast exposed to DOX, MBQ-167 or in combination. The maximum growth, doubling time and budding polarity of recombinant yeast with the Tet-Off system were reduced when treated with DOX (EC50=10ug/mL), as expected. The same effect was seen in the group treated with MBQ-167 (ED50=100uM), both alone and in combination with DOX. Ultimately, pull-down assays of the active Cdc42-GTP complex confirmed our hypothesis that MBQ-167 disrupts cell polarity through impeding Cdc42 activation. Here we validated MBQ-167 as a Cdc42 inhibitor in another biologic context. and present a method for screening Cdc42 inhibitors and potential therapeutics against metastatic cancer.

Citation Format: Michael John Rivera-Robles, Julia Medina-Velázquez, Gabriela M. Asencio-Torres, Sahily González-Crespo, Brian C. Rymond, José Rodríguez-Medina, Suranganie Dharmawardhane. Anti-metastatic drug MBQ-167 regulates cell polarity via Cdc42 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3994.
Abstract 4000: Hepatitis B virus X gene accelerates the development of HBV-related hepatocellular carcinoma
Authors: ZHANG Yu , YAN Qian , YU Dandan , GUAN Xinyuan
Published: August 2018

Epidemic studies have shown hepatocellular carcinoma (HCC) is highly associated with chronic HBV infection. However, the underlying mechanism of how HBV integration leading to HCC remains elusive. We applied the ViralFusionSeq algorithm to aid our analysis of the raw whole-genome sequencing data from TCGA database and found HBX “re-integration” in tumor cells showed a relatively consistent pattern compared with the adjacent normal liver tissue. In order to further elucidate the oncogenic role of truncated HBX gene in HCC, full-length and two COOH-terminal truncated HBX (X120 and X134) genes were stably transfected into HCC cell lines to investigate its function. Results showed that both truncated forms (X120 and X134) overexpression increased HCC cell proliferation rate, the ability of foci formation and colony formation in soft agar, as well as migration and invasion, while full-length HBX showed the opposite effect, both in vitro and vivo. Furthermore, RNA-sequencing of HBX overexpression HCC cells revealed key cancer-related signaling pathways were altered compared with control cells. Here our findings give a novel insight into the HBV integration pattern in the host genome which contributes to HCC.

Citation Format: Yu ZHANG, Qian YAN, Dandan YU, Xinyuan GUAN. Hepatitis B virus X gene accelerates the development of HBV-related hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4000.