bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2138: Complex copy number profiles in oligodendroglioma associate with anaplastic morphology and recurrence
Authors: Lowe Steve , Lindhorst Scott , Cachia David , Olar Adriana
Published: August 2019

Oligodendroglioma (O) comprises approximately 6% of all diffuse gliomas (DG) and is associated with favorable outcome. It is characterized by the presence of 1p/19q full arm co-deletion and concurrent IDH mutations. Despite a general favorable outcome selected patients with Os experience repeated radiological and histological progression. Copy number aberrations in Os may highlight profiles that correlate with patient's outcome.

Thirty-three Os from the files of our institution were profiled by immunohistochemistry, sequencing, and SNP-microarray. Pertinent clinical, pathological, and radiological data was collected. Descriptive statistics and survival analyses were performed.

Median age at initial diagnosis was 39 (range:19-80 years). Male to female ratio=1.2. All tumors were supratentorial with most common location the frontal lobe (21/33, 64%). Six tumors were anaplastic (WHO grade III). Nine patients experienced tumor recurrence, of which 5 experienced multiple recurrences. Only one patient died after 9.8 years from initial diagnosis. The median follow-up time = 2.5 years (range:0.01-23.6). 21/33(63%), 9/33(27%), and 3/33(0.09%) underwent subtotal, gross total resection, or biopsy respectively. 48%(16/33) patients received chemotherapy and/or radiation therapy at initial diagnosis. Most tumors (28; 85%) had additional copy number aberrations most commonly involving chromosomes 11q, 12q, 9p, 4q, 3p, 11p. Chromosomal deletions/full-arm losses were more common than gains. Diploidy/tetraploidy was detected in 2 cases and chromothripsis in 1. Anaplastic and recurrent tumors had more complex copy numbers aberrations, including numerous loci of loss of heterozygosity. Statistical significance however was not reached. Additional copy number aberrations characterize Os. Complex copy number profiles associate with aggressive morphology and recurrence.

Citation Format: Steve Lowe, Scott Lindhorst, David Cachia, Adriana Olar. Complex copy number profiles in oligodendroglioma associate with anaplastic morphology and recurrence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2138.
Abstract 2147: Galectin-1: A marker for claudin-low breast cancer
Authors: Balestrieri Kassondra , McDaniel Moses , Shoopman Christiana , Vohra Nasreen , Verbanac Kathryn
Published: August 2019

Metastasis and mortality among patients with triple negative breast cancer (TNBC) of the claudin-low subtype remain high because these tumors are clinically aggressive, chemoresistant, and lack targeted therapies. We have previously used proteomic approaches to analyze orthotopic and metastatic tumors from the syngeneic murine T11 tumor model, which displays gene expression profiles that mirror human claudin-low TNBC. These studies identified Galectin-1 (Gal-1), an N-acetyllactosamine-binding protein, as an overexpressed protein in cultured T11 cells, primary tumors, and lung metastases compared to normal lung and mammary fat pad tissue from healthy control mice. These results were confirmed by Western immunoblot and immunohistochemistry and were further supported by analysis of murine tumor microarray data sets, which revealed Gal-1 overexpression in the claudin-low subtype compared to other breast cancer subtypes. Here we extend our murine studies to investigate Gal-1 expression in breast cancer patients, with specific inclusion of claudin-low tumors. To compare differential Gal-1 expression across breast cancer subtypes, we initially analyzed published human microarray data sets from normal breast and primary breast tumors (total n=423, of which 51 are the claudin-low subtype): the highest Gal-1 expression was observed in claudin-low breast cancer. In order to validate these findings, we are examining breast cancer specimens from a unique cohort of ethnically diverse patients from rural Eastern North Carolina with a median follow-up of over 8 years. RNASeq data from a subset of this cohort (n=121) was used to assign subtypes (using PAM50 and the claudin-low classifier). Further studies are in progress to compare Gal-1 expression between subtypes and correlate with clinicopathologic factors and outcome. Immunohistochemical Gal-1 staining of primary tumor tissue blocks is underway to confirm expression in each subtype at the protein level. Immunohistochemistry will also define Gal-1 expression in the cell types comprising the heterogeneous tumor microenvironment (tumor, stromal, immune cells) and its subcellular localization (nuclear, cytoplasmic, membrane-associated). Through this ongoing work evaluating Gal-1 across breast cancer subtypes, we will help corroborate initial findings from the T11 claudin-low mouse model and yield further insights into the role of Gal-1 in breast cancer progression.

Citation Format: Kassondra Balestrieri, Moses McDaniel, Christiana Shoopman, Nasreen Vohra, Kathryn Verbanac. Galectin-1: A marker for claudin-low breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2147.
Abstract 2154: A novel dual-function polymeric scaffold for cancer drug screening
Authors: Wu Tianfu , Lin Zuan-Tao
Published: August 2019

Cell-based assay has been used as a conventional strategy for discovery of new anti-cancer drugs in pharmaceutical industries. Traditionally, two-dimensional (2D) cell culture based cytotoxicity assays are the most common approaches, despite the fact that they have significant limitations in simulating tissue or organ microenvironment. In comparison, 3-dimensional (3D) cell culture provides a more promising tool for new drug discovery, however, next-generation cell growth monitoring device for 3D cell culture is still challenging. Recent studies have shown the potential of metallic nanoparticle-polymeric composites in biosensing systems. We hypothesize that in a polymer-based 3D-cell culture system, cancer cell growth and the subsequent spheroid formation can cause structural change of the polymer network. With the incorporation of metallic nanoparticles into the polymeric network, electrical conductance change of the 3D-scafold may be monitored to reflect cell growth and spheroid formation. Here, we developed a dual-function polymer scaffold (DPS) by integrating thermosensitive polymers, silver flake-decorated polymers, and gNP-decorated polymers as a biosensor to monitor cancer cell growth. We found that the Triple negative breast cancer stem cell line MDA-MB-231 formed spheroid in DPS. It is notable that the increase of cancer cell numbers during the formation of spheroid was proportional to the change of electrical conductance of the 3D-scafold. Importantly, the results from DPS cell growth monitoring method correlate well with conventional cell counting during 3D cell culture of breast cancer cells. SEM imaging and AFM imaging have confirmed the integration of silver flakes and gold nanoparticles in the DPS. Finally, we demonstrated that the DPS 3D-scafold exhibited robust responses to MDA-MB-436 cells when exposed to 3 nM of paclitaxel (PTX) and docetaxel (DTX), respectively. This dual-function 3D-scaffold, for both cell growth and growth monitoring may have a good potential in drug screening in the future.

Note: This abstract was not presented at the meeting.

Citation Format: Tianfu Wu, Zuan-Tao Lin. A novel dual-function polymeric scaffold for cancer drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2154.
Abstract 2164: In-licensing and knockout validation of monoclonal hybridomas to improve antibody reproducibility
Authors: Heaton Sam , Koch Sofia , Ge Jiangyan , Rizzo Joshua , Prater Michael , Dreja Hanna , Hamilton Bruce , Solache Alejandra
Published: August 2019

Antibodies are the most common tools in basic science and clinical research. Over the past several years there has been an ongoing discussion around antibody validation in light of poor experimental reproducibility between laboratories. This is in part due to the cross-reactivity between antibodies and off-target proteins and the variability between different antibody batches. The resulting experimental irreproducibility leads to wasted resources and compromises the advancement of science. Abcam partners with various research institutes and biotech companies to in-license hybridoma cell lines producing monoclonal antibodies. Through this in-licensing program, Abcam has taken steps to help ensure target-specificity of valuable clones. Our in-licensing program utilizes our ISO9001-accredited production method to ensure each batch performs in-line with previous batches. Where possible, all in-licensed antibodies are also tested in knockout (KO) material to confirm the specificity of the in-licensed clone and subsequent batches. Through our partnership with Horizon Discovery to test antibodies with KO cells, target genes are mutated via CRISPR-Cas9 within a haploid cell line, which results in a frameshift and a complete loss of gene expression. KO cell lines provide a true negative control for antibody validation that allows us to confirm the specificity of clones we in-license and produce. Alongside KO, we are making use of our expertise in recombinant antibodies to explore the conversion of selected in-licensed hybridomas to recombinant antibodies. A key issue with monoclonals is degradation of the clone over time, leading to inconsistent batches and irreproducible results. By sequencing and cloning the antibody, we expect to guarantee continued specificity and reproducibility; therefore, ensuring our licensors see long term impact and value generation for their innovations. Here we present KO data for several in-licensed published clones, validated in both western blot and immunocytochemistry. The antibodies have been tested in KO and wild-type cells to confirm their specificity. By providing researchers with reliable and specific antibodies that work as intended, we hope to minimize wasted resources and improve reproducibility. In addition to our KO data, we present data from the recombinant conversion of clone NAT105 to PD1 and show comparable, if not superior, performance when compared with the hybridoma.

Citation Format: Sam Heaton, Sofia Koch, Jiangyan Ge, Joshua Rizzo, Michael Prater, Hanna Dreja, Bruce Hamilton, Alejandra Solache. In-licensing and knockout validation of monoclonal hybridomas to improve antibody reproducibility [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2164.
Abstract 2167: Antibody glycoengineering for drug delivery applications
Authors: Sehgal Drishti Navin , Kalscheuer Stephen , Panyam Jayanth
Published: August 2019

Antibody-drug conjugates (ADC) hold considerable promise as anticancer agents. A critical determinant of the effectiveness of ADCs is the chemistry that is used to conjugate the payload. Currently used approaches include primarily conjugation to either side-chain amine or carboxylic acid groups or conjugation to thiols. Because these reactions are not site specific and not easily controlled, these chemistries can result in reduced affinity for the target antigen. Further, these conjugation reactions lack selectivity and can result in heterogeneous mixtures of products that differ in the sites and stoichiometry of modification. We investigated a glycoengineering strategy that enables the introduction of artificial azide groups in the antibody without affecting their antigen affinity. This is based on the observation that glycosyltransferases can incorporate non-natural sugars (e.g., azido mannose) at different sites on an IgG molecule. The azide groups in these artificial sugars are then available to react with alkynes through copper-catalyzed ‘click’ chemistry or with strained alkynes such as dibenzyl cyclooctyne (DBCO) allowing for biorthogonal, copper-free ‘click’ chemistry. Because the sugars are added reproducibly and at a site that does not affect antigen binding, the glycoengineering technology would overcome problems associated with traditional conjugation strategies. Using this approach, azide groups were introduced in anti-CD133 and anti-perlecan antibodies. Further, the azide groups were available to react with various DBCO conjugates including fluorophores, drug molecules and nanoparticles. Importantly, the addition of artificial sugar and subsequent azide-alkyne reaction did not affect the affinity of the antibody for the target antigen. Conjugation of a cytotoxin to the antibody resulted in enhanced cell kill in vitro and efficacy in vivo. We plan on further improving the effectiveness of this approach by investigating cleavable linker technology in combination with the glycoengineering strategy. We expect that this strategy will prove to be a unique platform technology that will have a significant impact on antibody-based therapeutics.

Citation Format: Drishti Navin Sehgal, Stephen Kalscheuer, Jayanth Panyam. Antibody glycoengineering for drug delivery applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2167.
Abstract 2179: Antitumor activity of JNJ-63576253 (TRC253), a small molecule antagonist of F877L mutant and wild-type androgen receptor
Authors: Bush Tammy L. , Ndikuyeze Georges Habineza , Bignan Gilles , Branch Jonathan , Ondrus Janine , Shi Yifan , Luistro Leopoldo , Hastings James , Erhardt Joseph , Hickson Ian , Patel Shefali , Connolly Peter , Zhang Zhuming , Bischoff James , Rupnow Brent , Gottardis Marco , Packman Kathryn
Published: August 2019

Androgen receptor (AR) antagonists have transformed prostate cancer patient care by targeting a key nodal point in tumor cell signaling. However, despite the impressive clinical activity of first- and second-generation antiandrogens, acquired resistance frequently emerges. Point mutations in the ligand-binding domain of AR, such as phenylalanine to leucine at position 877 (ARF877L), account for 10-20% of resistance. Such mutations are characterized by receptor activation, rather than inhibition, by first- and second-generation antiandrogen therapeutics.

JNJ-63576253 is a potent, high affinity competitive binder of wild type and mutant AR, including F877L. JNJ-63576253 blocks AR nuclear translocation, AR binding to DNA, and AR-dependent transcription. JNJ-63576253 inhibits the proliferation of androgen receptor driven prostate cancer cell lines, including those bearing ARF877L.

In the Hershberger assay in male Sprague Dawley rats, oral administration of JNJ-63576253 inhibited androgen sensitive organ (ASO) development in a dose-dependent manner. In male SHO mice bearing LNCaP xenografts with either wild-type or ARF877L, daily treatment with 30 mg/kg JNJ-63576253 treatment resulted in statistically significant antitumor activity, whereas second-generation antiandrogen enzalutamide had no antitumor efficacy in the LNCaP ARF877L mutant model.

Janssen and Tracon Pharma have entered a strategic licensing collaboration, whereby Tracon possesses exclusive rights for clinical development of JNJ-63576253 (now called TRC253). Tracon has entered TRC253 into Ph1/2A clinical evaluation in metastatic castration-resistant prostate cancer patients.

Citation Format: Tammy L. Bush, Georges Habineza Ndikuyeze, Gilles Bignan, Jonathan Branch, Janine Ondrus, Yifan Shi, Leopoldo Luistro, James Hastings, Joseph Erhardt, Ian Hickson, Shefali Patel, Peter Connolly, Zhuming Zhang, James Bischoff, Brent Rupnow, Marco Gottardis, Kathryn Packman. Antitumor activity of JNJ-63576253 (TRC253), a small molecule antagonist of F877L mutant and wild-type androgen receptor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2179.
Abstract 2181: A novel CXCR4 antagonist strongly mobilizes hematopoietic stem cells in vivo
Authors: Fang Xiao , Meng Qian , Fang Xiong , Mao Yujia , Xu Yan , An Jing , Huang Ziwei
Published: August 2019

Introduction: The transplantation of hematopoietic stem cells (HSCs) has been the standard of care for the treatment of chemotherapy sensitive relapsed acute leukemia. HSCs mainly reside in the bone marrow with only few HSCs circulating in the peripheral blood (PB), due to homing effect mediated by the interaction of chemokine receptor CXCR4 expressed on HSCs with chemotactic ligand SDF-1α rich in the bone marrow. It is clinically desirable to enhance the number of HSCs in the PB for collection during the transplantation procedure. Since SDF-1α/CXCR4 axis and interaction mediates the homing and retention of HSCs in the bone marrow, our laboratory has recently discovered and developed a novel small molecule, HFX51116, that is capable of binding CXCR4 and blocking its interaction with SDF-1α, raising the possibility that this compound can be used for releasing HSCs from the bone marrow to the PB useful for the transplantation of HSCs.

Method and Results: In vitro, there was almost no toxicity of HFX51116 at high concentrations in bone marrow stromal cells that were originally extracted from rats. In addition, HFX51116 was stable in rat serum. In vivo, we tested the short-term and long-term mobilization efficacy of HFX51116. For the short-term efficacy study, we determined the dynamic change of efficacy at different times and doses of HFX51116 in HSC mobilization assays. Furthermore, different strains of mice, such as C57/BL6, C3H/HEJ and DBA/2, were sensitive to HFX51116. HFX51116, when used in combination with G-CSF, had higher efficacy than G-CSF alone. For the long-term repopulation study, we employed the CD45.1/CD45.2 competitive assay to demonstrate that the HSCs mobilized by G-CSF and HFX51116 had the repopulation ability when compared with the bone marrow cells.

Conclusion: Our studies have demonstrated high HSC mobilization efficacy of HFX51116 in vivo and the promise of this new lead compound for further preclinical and clinical studies.

Citation Format: Xiao Fang, Qian Meng, Xiong Fang, Yujia Mao, Yan Xu, Jing An, Ziwei Huang. A novel CXCR4 antagonist strongly mobilizes hematopoietic stem cells in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2181.
Abstract 2192: Concurrent Tumor Treating Fields (TTFields) and dexamethasone decrease proliferation and clonogenicity of patient-derived glioblastoma cells in vitro
Authors: Michelhaugh Sharon K. , Kiousis Sam , Mittal Sandeep
Published: August 2019

Background: TTFields are FDA-approved for treatment of newly-diagnosed and recurrent glioblastoma and extend progression-free survival and overall survival in newly-diagnosed glioblastoma patients. Dexamethasone is administered perioperatively to GBM patients to reduce tumor-associated edema and increased intracranial pressure. In addition, high dose corticosteroids may be required during TTFields therapy. In this study, a patient-derived GBM cell line from a newly-diagnosed glioblastoma patient was treated in vitro with TTFields ± dexamethasone followed by proliferation and clonogenic assays.

Methods: This study was approved by the Institutional Review Board and written consent obtained from the patient. A single-cell suspension was generated from tumor tissues obtained from the newly-diagnosed patient. Cells were cultured in DMEM/F12 media with 10% fetal bovine serum and gentamicin. Prior to TTFields initiation, cells were plated on plastic coverslips (1×104cells/coverslip; n=7-8/group) and incubated overnight. Then, TTFields were applied at 200 kHz with a field intensity of ~1.6 V/cm for 14 days. Cells were treated with TTFields with or without 50 µM dexamethasone. At the end of the TTFields application, proliferation was assessed with XTT assay and cells were harvested and replated for the clonogenic assay (n=3/group). XTT data and clonogenic assay cell counts were analyzed by one-way ANOVA with Tukey’s post-hoc test.

Results: In patient-derived glioblastoma cells treated with TTFields and dexamethasone for 14 days, proliferation was significantly decreased compared to the dexamethasone-only and control groups (0.483±0.30; 1.33±0.18; 1.42±0.11, respectively, ANOVA p<0.001). Similarly, cell counts in the clonogenic assay were markedly decreased in the TTFields ± dexamethasone group compared to dexamethasone-only (1184±425.9; 46464±4580; p<0.05).

Conclusions: In glioblastoma cells from a newly-diagnosed patient, concurrent TTFields with dexamethasone produced a ~three-fold reduction of proliferation and reduced clonogenicity compared to control groups.

Citation Format: Sharon K. Michelhaugh, Sam Kiousis, Sandeep Mittal. Concurrent Tumor Treating Fields (TTFields) and dexamethasone decrease proliferation and clonogenicity of patient-derived glioblastoma cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2192.
Abstract 2193: AXL inhibitor TP-0903 attenuates AXL-TGFbeta Hippo signaling axis in lung adenocarcinoma cells
Authors: Taverna Josephine Amalia , Hung Chia-Nung , Lin Chun-Lin , Osmulski Pawel , Chen Meizhen , Wang Chiou-Miin , Lucio Nicholas L. L. , Kirma Nameer , Chou Chih-Wei , Gaczynska Maria E. , Nazarullah Alia , Wade Mark , Mouritsen Lars , Huang Tim
Published: August 2019

Background: Non-small cell lung cancer (NSCLC) is a molecularly heterogeneous disease with a high propensity for drug resistance and metastasis. AXL, a member of the Tyro3-AXL-Mer family of receptor tyrosine kinases, is a central regulator of epithelial-to-mesenchymal transition (EMT) and enables tumor cells to invade and acquire drug resistance. AXL is overexpressed in lung tumors, correlates positively with tumor invasion, drug resistance, and negatively predicts overall survival. We mechanistically interrogate the effects of AXL inhibitor TP-0903 on EMT in lung adenocarcinoma cells using transcriptomic and proteomic profiling.

Methods: Atomic force microscopy, Western blot analysis, RNA sequencing and mass cytometry (CyTOF) were all used to scrutinize the biomechanical properties, phenotypic, transcriptomic and proteomic profiles of A549 cells treated with 40nM TP0903 or shAXL knockdown.

Results: TP-0903 attenuates total AXL/AXL phosphorylation and blunts transcriptional responses to TGFβ-Hippo signaling by disrupting the transcriptional complexes formed by SMAD2/3, SMAD4, YAP1 and TAZ. AXL knockdown or TP-0903 reverses EMT phenotype and reduces migration potential in A549 and H2009 adenocarcinoma cell lines. CyTOF data also identified resistant clones that overexpress TGFβ receptor II, TAZ protein and display hybrid EMT phenotypes.

Conclusions: We are the first to report the interplay between AXL and TGFβ-Hippo signaling axis. TP-0903 study agent has excellent therapeutic promise in NSCLC and we speculate that TP-0903 drug can target epithelial to mesenchymal transitional states in lung cancer cells possibly through the inhibition of the AXL-TGFβ-Hippo signaling axis.

Citation Format: Josephine Amalia Taverna, Chia-Nung Hung, Chun-Lin Lin, Pawel Osmulski, Meizhen Chen, Chiou-Miin Wang, Nicholas L. L. Lucio, Nameer Kirma, Chih-Wei Chou, Maria E. Gaczynska, Alia Nazarullah, Mark Wade, Lars Mouritsen, Tim Huang. AXL inhibitor TP-0903 attenuates AXL-TGFbeta Hippo signaling axis in lung adenocarcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2193.
Abstract 22: Design and synthesis of substituted tetrahydroisoquinoline derivatives as anti-angiogenic and anti-breast cancer agents
Authors: Redda Kinfe Ken , Gangapuram Madhavi , Eyuinni Suresh
Published: August 2019

Angiogenesis is the formation of new blood vessels and plays a critical role in the development, invasion and metastasis of breast cancer pathogenesis. Vascular Endothelial Growth Factor (VEGF) is one of the important angiogenic factors secreted by tumor cells. Overexpression of VEGF significantly increases tumor growth and angiogenesis in a murine model of breast cancer. Inhibition of VGEF pathway by limiting new blood vessels formation has proven to be a valuable approach in reducing tumor growth. During the last two decades, diversified small molecules were evaluated and developed as anti-angiogenic agents. Trabectedin (ET-743; Yondelis) is a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate Ecteinascidia turbinata and has a potent in vitro and in vivo antitumor activity, including prostate and breast cancers. Trabectedin significantly inhibited the mRNA and protein expression levels of VEGF in all breast cancer cells tested. Previously, we developed a series of tetrahydroisoquinoline (THIQs) derivatives and their in-vitro anti-proliferative activities were tested against MCF-7, MDA-MB-231 human breast cancer cell lines and Ishikawa human endometrial adenocarcinoma cell lines. These THIQ derivatives have the same core structure as that of Trabectedin alkaloid and are considered potential anti-angiogenic agents. Novel substituted THIQ derivatives were designed and synthesized in our laboratory. N-amination of substituted isoquinolines by the aminating agent, O-mesytelenesulfonylhydroxylamine and treated with acyl chlorides led to the formation of ylides. The ylides were reduced using sodium borohydride to yield the desired substituted tetrahydroisoquinolines in moderate to good yields. The synthesized compounds were characterized using NMR and elemental analysis. Of the synthesized compounds, fifteen compounds were screened for anti-angiogenesis activity in the Eli Lilly’s Open Innovation Drug Discovery Program (OIDD). Five compounds exhibited greater than 50% inhibition of the tube area at 10µM in the primary single point assay. These five compounds were further screened at different concentrations to obtain dose response curves and IC50 Values. Compound 4-ethyl-N-(3,4-dihydroisoquinolin-2(1H)-yl) benzamide showed an IC50 Value of 1.72 µM.

This research was supported by the National Center for Research Resources and the National Institute of Minority Health and Health Disparities of the National Institutes of Health through Grant Number 8 G12MD007582-28.

Citation Format: Kinfe Ken Redda, Madhavi Gangapuram, Suresh Eyuinni. Design and synthesis of substituted tetrahydroisoquinoline derivatives as anti-angiogenic and anti-breast cancer agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 22.
Abstract 2207: The dual role of BI 2536, a small-molecule inhibitor that targets PLK1, in induction of apoptosis and attenuation of autophagy in neuroblastoma cells
Authors: Li Zhiheng
Published: August 2019

Neuroblastoma(NB) is the most common extra-cranial solid tumor in childhood with the overall survival less than 40%. Polo-like kinase 1 (PLK1) is a serine/threonine-protein kinase expressed during mitosis and overexpressed in multiple cancers, including neuroblastoma. We found the PLK1 expression related to the outcome of NB patients. BI2536, a small molecule inhibitor against PLK1, significantly reduced cell viability in a panal of NB cell lines, with IC50 less than 100 nM. PLK1 inhibition by BI 2536 treatment induced cell cycle arrest at G2/M phase and cell apoptosis in NB cells. Realtime PCR array revealed the PLK1-regulated genes, such as BIRC7, TNFSF10, LGALS1 and DAD1 et al. Moreover, autophagy activity was investigated in the NB cells treated with BI 2536. BI 2536 treatment in NB cells increased LC3-II puncta formation and LC3-II expression. Formation of autophagosome induced by BI 2536 was observed by TEM. However, BI2536 abrogated the autophagic flux in NB cells by reducing SQSTM1/p62 expression and AMPKαT172 phosphorylation. These results provide new clues for the molecular mechanism of cell death induced by BI 2536 and suggest that BI 2536 may act as new candidate drug for neuroblastoma.

Citation Format: Zhiheng Li. The dual role of BI 2536, a small-molecule inhibitor that targets PLK1, in induction of apoptosis and attenuation of autophagy in neuroblastoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2207.
Abstract 2214: Development and validation of PredicineATLAS, a 600-gene liquid biopsy panel for simultaneous detection of TMB and genomic alterations in Immuno-Oncology
Authors: Zhao Zhixin , Chang Amy , Cheung Tak , Xie Feng , Montesinos Carlos , Wang Amy Xiaohong , Zhou Kemin , Jia Shidong , Yu JIanjun , Du Pan
Published: August 2019

Introduction: Tumor mutation burden (TMB) is a measurement of the number of somatic mutations carried by tumor cells. Microsatellite instability (MSI) reflects the genetic hypermutability. Both TMB and MSI have been shown to be effective genomic biomarkers for predicting response to anti PD-1/PD-L1 immunotherapy. Whole exome sequencing (WES) is a traditional means to estimate TMB and MSI from tumor tissue, however, it is challenging to translate it into blood or urine due to its high expense and low tumor cell percentage. In the present study, we developed a 600-gene liquid biopsy TMB panel, PredicineATLAS, to measure genomic alterations and estimate TMB and MSI using human body fluids.

Methods: The PredicineATLAS panel contains 600 genes and covers 2.4Mb genomic regions (1.4 Mb of exonic coding regions). It was specifically designed for immuno-oncology, covering comprehensive immune- and cancer-related genes, biomarkers and MSI sites that are associated with anti PD-1/L1 therapy response. Reference materials with known genomic alterations and tumor cell lines were titrated with different concentrations to mimic different tumor contents for TMB and MSI measurement. Matched tissue and blood were used in validation study.

Results: PredicineATLAS panel-based TMB strongly correlated with the WES-TMB using TCGA dataset. Analytical validation showed that PredicineATLAS NGS assay is highly robust and sensitive for detection of genomic alterations. Using matched tissue and blood sample set, blood-based PredicineATLAS panel-TMB and MSI correlate with its counterpart derived from tissue WES.

Conclusions: PredicineATLAS liquid biopsy assay was developed and validated using reference materials and clinical samples. It detects genomic alterations and accurately estimate TMB and MSI, providing valuable genomic information to guide immuno-oncology therapy and clinical drug development.

Citation Format: Zhixin Zhao, Amy Chang, Tak Cheung, Feng Xie, Carlos Montesinos, Amy Xiaohong Wang, Kemin Zhou, Shidong Jia, JIanjun Yu, Pan Du. Development and validation of PredicineATLAS, a 600-gene liquid biopsy panel for simultaneous detection of TMB and genomic alterations in Immuno-Oncology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2214.
Abstract 2217: Serum metabolic profiling identified a distinct metabolic signature in bladder cancer smokers: A key metabolic enzymes associated with patient survival
Authors: Amara Chandra S. , Ambati Chandrashekar R. , Vantaku Venkatrao , Danthasinghe Piyarathna W b , Donepudi Sri Ramya , Ravi Shiva S. , Arnold James M. , Putluri Vasantha , Chatta Gurkamal , Guru Khurshid A. , Badr Hoda , Terris Martha K. , Bollag Roni , Sreekumar Arun , Apolo Andrea B. , Putluri Nagireddy
Published: August 2019

Urinary bladder cancer (BLCA) is the 9th most common malignant disease and the 13th most common cause of cancer death worldwide. Occupational exposure to carcinogens has been long associated with increased BLCA risk. Tobacco smoke contains more than 60 carcinogens causing at least 18 different types of cancer including BLCA. Reliable biomarkers for accurately predicting survival in Bladdder cancer (BLCA) smokers is lacking due to complex genomic and transcriptomic heterogeneities associated with the disease. We performed liquid chromatography-mass spectrometry (LC-MS) based targeted metabolomic analysis for >300 metabolites in serum obtained from two independent cohorts of BLCA never smokers, smokers, healthy smokers, and healthy never smokers. 40 metabolites (FDR <0.25) were identified to be differential between BLCA never smokers and smokers. Increased abundance of amino acids (tyrosine, phenylalanine, proline, serine, valine, isoleucine, glycine, asparagine) and taurine levels were observed in BLCA smokers. Integration of differential metabolomic gene signature and transcriptomics from TCGA cohort resulted in intersecting 17 gene signature that showed significant correlation with patient survival in BLCA smokers. Importantly Catechol-O-Methyltransferase (COMT), Iodotyrosine Deiodinase (IYD), and Tubulin Tyrosine Ligase (TTL) showed a significant association with patient survival in publicly available BLCA smokers datasets and did not have any clinical association in never smokers.

Note: This abstract was not presented at the meeting.

Citation Format: Chandra S. Amara, Chandrashekar R. Ambati, Venkatrao Vantaku, Piyarathna W b Danthasinghe, Sri Ramya Donepudi, Shiva S. Ravi, James M. Arnold, Vasantha Putluri, Gurkamal Chatta, Khurshid A. Guru, Hoda Badr, Martha K. Terris, Roni Bollag, Arun Sreekumar, Andrea B. Apolo, Nagireddy Putluri. Serum metabolic profiling identified a distinct metabolic signature in bladder cancer smokers: A key metabolic enzymes associated with patient survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2217.
Abstract 2218: Exosomal PD-L1 and T lymphocyte status predict the effect of anti-PD-1 therapy
Authors: Qu Xiujuan , Zhang Chaoxu , Fan Yibo , Che Xiaofang , Liu Yunpeng
Published: August 2019

Background: Anti-PD-1 therapy is approved to be remarkable for cancer treatment. However, the response rate is dissatisfactory. PD-L1 expression in tumor tissue, which affected by chemotherapy, radiotherapy and activity of signal transduction pathways, is unreliable to predict treatment response. Recent studies suggest that exosomal PD-L1 not only exerts immunosuppressive effects besides tumors, but also plays a significant role in development of tumor microenvironment in the distance. A deeper research of exosomal PD-L1 assessment is needed to improve the predictive value and treatment efficacy.

Method: We enrolled 28 patients with advanced tumor, including 8 lung adenocarcinoma, 7 lung squamous carcinoma, 4 esophageal carcinoma, 2 colorectal carcinoma, 2 cholangiocarcinoma, 1 small cell lung cancer, 1 gastric carcinoma, 1 duodenal adenocarcinoma, 1 nasopharyngeal carcinoma and 1 tongue squamous cell carcinoma, who were treated by anti-PD-1 therapy followed the standard regimens. Exosomes were collected and purified from plasma with exosome isolation kit. Exsomal PD-L1 was detected by ELISA. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll gradient. Exhausted T lymphocyte were stained for cytometry.

Result: Compared with responder, exosomal PD-L1 of non-responders is significantly higher than responder (P=0.036) before treatment. Moreover, the CD4+ and CD8+ T lymphocyte count is higher and the percentage of exhausted T lymphocyte rate is lower in responder than the non-responder cohort before treatment.

Although there is no statistical difference, the level of exosomal PD-L1 decreased in responder cohortat on 6 to 8 weeks after treatment, on the contrary, increased in non-responder cohort after anti-PD-1 therapy. Concurrently, exosomal PD-L1 and tumor burden were decreased when the therapy was effective.

Conclusion: Low levels of exosomal PD-L1 and exhausted T lymphocyte rate on baseline and decreasing expression of exosomal PD-L1 after treatment could screen out the potential benefit populations of anti-PD-1 therapy in a variety of solid tumors. These findings suggest that expression of exosomal PD-L1 and exhausted T lymphocyte cell rate could be attractive and predictive biomarkers for clinical responses to anti-PD-1 treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Xiujuan Qu, Chaoxu Zhang, Yibo Fan, Xiaofang Che, Yunpeng Liu. Exosomal PD-L1 and T lymphocyte status predict the effect of anti-PD-1 therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2218.
Abstract 2222: Progastrin a new biomarker for hepatocellular cancer patient follow-up
Authors: Prieur Alexandre , Dupuy Marie , Joubert Dominique , Assenat Eric
Published: August 2019

Background and aims: Alpha-fetoprotein (AFP) is the most widely used biomarker for hepatocellular carcinoma (HCC) patient follow-up, even though around 30% of HCC do not express AFP and its capacity to reliably monitor treatment efficacy is questioned. Interestingly, progastrin, a direct target of the oncogenic Wnt/beta-catenin pathway that is commonly activated in HCC, is secreted as such from tumor cells of various origin, from the very first steps of tumorigenesis. Here, we show the first study evaluating the potency of blood progastrin levels to monitor treatment efficacy and compared it with AFP.

Methods: Progastrin was quantified in the blood of 87 patients with HCC (focal (n=23), locally advanced (n=49) or metastatic (n=19). AFP was available in 79 patients. The cancerREAD technology (CE marked since 2017) specifically designed to detect progastrin with a high sensitivity and not any maturation products such as gastrin or gastrin glycine extended was used to assay progastrin in the blood. Patients received a systemic (nevaxar, tepotinib, regorafenib, nivolumab, anti FGR, or carbozantinib) and/or a local treatment. Variations in the levels of progastrin and AFP were analyzed in relationship with evolution of the disease: progression, decrease, stable, in remission, without taking into account the different treatment regimens.

Results: Progastrin was detected in 80% of the patients (cut-off value 1 pM, range 0 to 75.9 pM, mean value +/- SE 16.29 +/-1.97 pM). Patients in remission had a significantly lower blood progastrin compared to focal (p=0.019), locally advanced (p=0.0001) or metastatic HCC (p=0.015) (Focal 16.96 +/- 4.82 pM; locally advanced 18.57 +/- 2.75 pM; metastatic 17.81 +/- 4.9 pM; in remission 4.89 +/- 1.05 pM). AFP was above 10 ng/ml (a common threshold value) in 59.4% of the samples; levels of progastrin were not statistically correlated with those of AFP. Blood samples for follow-up of treatment were available for 48 patients. Compared to AFP, progastrin was a better biomarker of the evolution of the disease in 35.1% of the patients. Both biomarkers were equivalent in 32.4% and none were good in 10.8%. AFP was better than progastrin only in 21.6%.

Conclusion: Thus, and for the first time, we showed that not only progastrin is a better blood biomarker than AFP to detect HCC but also is better for the follow-up of HCC patients, whatever the treatment of the patient. This is the first study that shows the potential of progastrin (alone or in combination with AFP) as a tool for follow-up of cancer treatment efficacy.

Citation Format: Alexandre Prieur, Marie Dupuy, Dominique Joubert, Eric Assenat. Progastrin a new biomarker for hepatocellular cancer patient follow-up [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2222.
Abstract 2225: Exosomes in plasma for protein based cancer biomarkers: A clinical research tool using UHPLC-MS
Authors: Sarracino David A. , Xie Xiaolei , Martel Courtney E. , Luan Shen , Rosenblatt Kevin
Published: August 2019

Introduction:

Exosomes have been used as a source of biomarkers in diagnostics for many diseases, including cancer, and are now being used as therapeutic delivery systems. To date, the most efficient method for isolating exosomes from serum or plasma samples has been to use a size exclusion, chromatography (SEC) based isolation. The conditions of the SEC separation can be modified to yield extracellular vesicles of different sizes and diagnostic value. Apoptotic bodies, 50-5000nm, microvesicles, 50-1000nm, and exosomes, 30-120nm, all have different diagnostic and targeted applications. Here, we show a routine and robust method for the isolation and monitoring of proteins in exosomes isolated from plasma samples. The methods and conditions used here are focused on rapidly providing a high yield of exosome vesicles in a monodisperse 40nm size range from a plasma sample.

Materials and methods:

K2EDTA plasma samples from donor disease and control groups were collected with IRB approval. All SEC columns and reagents for exosome isolation were obtained as a kit (NX Prenatal, Houston, TX). The 40nm exosome fraction from the column was lysed, the proteins reduced, alkylated and digested with trypsin; the subsequent peptides were analyzed by ultra-high performance liquid chromatography (Vanquish Horizon, ThermoFisher Scientific), tandem mass spectrometry (Q-Exactive, ThermoFisher Scientific), to quantify and identify greater than 500 proteins from exosomes, isolated from a single plasma sample, in a 1 hour method. Subsequent isolations of 40nm exosome fractions were labeled with a sulfo-NHS-biotin linker, followed by lysis and enrichment with streptavidin columns, and an identical LC-MS analysis to obtain quantitative analysis of the surface membrane protein contents on the exosomes.

Results:

Raw data was analyzed using the label-free quantitation module in Proteome Discoverer (Thermofisher Scientific). Peptide peak areas from three technical replicate runs were used to ascertain the reproducibility of the method across both normal and disease samples. Membrane protein extracts LC-MS data was further grouped by Protein Center (ThermoFisher Scientific) by cell type and tissue of origin.

Conclusions:

The methods shown here provide a robust and efficient platform for 40nm exosome isolation from human plasma samples, and quantitative analysis of the protein content thereof for the purposes of biomarker discovery, quantification and screening purposes.

Citation Format: David A. Sarracino, Xiaolei Xie, Courtney E. Martel, Shen Luan, Kevin Rosenblatt. Exosomes in plasma for protein based cancer biomarkers: A clinical research tool using UHPLC-MS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2225.
Abstract 223: Antibody intracellular activated drug conjugate (AiADC) as novel ADC technology
Authors: Suwa Atsushi , Komiya Kazuo , Kadotsuji Kenta , Ochiai Shoko , Takanashi Yosuke , Sugaru Eiji , Ban Hitoshi
Published: August 2019

Antibody drug conjugate (ADC) consisting of antibody and modifier (potent cytotoxic drug with chemical linker) is expected to selectively eliminate cancer cells expressing corresponding cell-surface antigen. More than 60 ADCs are in clinical trials and about 75 percent of its modifiers are based on three series (auristatin, maytansine, and pyrrolobenzodiazepine). Whereas some ADCs show promising clinical benefits, several ADCs may not have shown maximum value of ADC due to their narrow therapeutic windows. The function of modifiers impacts on total characteristics of ADC such as the efficacy, tolerability, and pharmacokinetics (PK). Therefore, further exploration of new types of modifier for ADC technology platform is needed. We designed hydrophilic modifiers which can be activated only inside of target cells. The ADC consisting of certain IgG1 monoclonal antibodies and this modifier (DSP-ADC) showed selective and potent cytotoxicity to target cells with IC50s at picomolar range. The DSP-ADC with the average drug-to-antibody ratio (DAR) of 7-8 was stable in blood circulation and showed favorable PK profiles in mice. Additionally, administration of the DSP-ADC led to dose-dependent suppression of cancer cell proliferation in mice xenograft models and achieved complete remissions at 3 mg/kg without body weight reduction. The toxicity studies of DSP-ADC in rats showed no effect on hematologic parameters up to 100 mg/kg. These modifiers are applicable to various monoclonal antibodies. Taken together, our modifiers can be fundamental component for ADC technology. We present here the novel ADC technology named as antibody intracellular activated drug conjugate (AiADC).

Citation Format: Atsushi Suwa, Kazuo Komiya, Kenta Kadotsuji, Shoko Ochiai, Yosuke Takanashi, Eiji Sugaru, Hitoshi Ban. Antibody intracellular activated drug conjugate (AiADC) as novel ADC technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 223.
Abstract 224: Antibody-drug conjugates (ADCs) of a new class of N-10 amino linked DNA alkylating indolino-benzodiazepines (IGNs)
Authors: Miller Michael L. , Reid Emily E. , Archer Katie E. , Harris Luke , Maloney Erin K. , Bartle Laura M. , Ab Olga , Wilhelm Alan J. , Ponte Jose F. , Singh Rajeeva , Keating Thomas A. , Chari Ravi V.
Published: August 2019

We have previously reported on our antibody-drug conjugates (ADCs) that incorporate a highly potent novel DNA alkylating indolino-benzodiazepine (termed IGN) dimer. ADCs of these DNA alkylating IGNs were found preclinically to demonstrate better tolerability and an improved overall therapeutic index (TI) compared with those of the corresponding DNA crosslinking IGNs (Miller, et al., Mol. Cancer Ther. 2016, 2018). IGN-containing ADCs that have reached the clinical stage bear DNA alkylating IGN compounds linked to the antibody via a cleavable linker, incorporated at the amino group of a centrally located anilino spacer moiety. In our ongoing effort to further explore the structure-activity relationship (SAR) of DNA alkylating effector molecules for ADCs, we disclose here a new class of IGNs possessing a self-immolative peptide linker attached at the N-10 amine of the imine-reduced IGN monomer subunit. We explored the impact of modifying the central spacer group connecting the IGN monomers, and site of linkage on in vitro potency and the interaction with DNA. A set of IGN molecules that met our potency criteria were identified and linkable forms of these compounds were prepared and conjugated to a folate receptor-α (FRα)-binding antibody. These ADCs displayed potent, antigen-specific in vitro activity across a panel of FRα-expressing cell lines. A lead ADC selected for in vivo studies demonstrated high stability in plasma and potent efficacy in various xenograft models, at doses well below the maximum tolerated dose. Thus, IGNs modified with an N-10 self-immolative peptide linker are promising DNA alkylating effector molecules for use in ADCs.

Citation Format: Michael L. Miller, Emily E. Reid, Katie E. Archer, Luke Harris, Erin K. Maloney, Laura M. Bartle, Olga Ab, Alan J. Wilhelm, Jose F. Ponte, Rajeeva Singh, Thomas A. Keating, Ravi V. Chari. Antibody-drug conjugates (ADCs) of a new class of N-10 amino linked DNA alkylating indolino-benzodiazepines (IGNs) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 224.