bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3880: Quantitative human secretome: a technology to measure the expression levels of thousand of secreted proteins
Authors: Huang Ruo-Pan , Mao Yingqing , Huang RuoChun , Yen Haw-Han
Published: July 2016

The secretome constitutes an increasingly important category of proteins which are involved in a multitude of biological, physiological and pathological processes, and are thus a clinically relevant source for biomarkers and therapeutic targets. The development of cancer often leads to alterations in growth-stimulatory autocrine and paracrine signals which change the expression levels of secreted proteins such as growth factors, cytokines, chemokines, and angiogenic factors. There is thus an urgent need to develop high-content and high-throughput approaches to profile the secretome. Using a sandwich-based ELISA format, we have developed an antibody array technology which can quantitatively measure expression levels of 1000 secreted proteins. Most of the target proteins are detectable at pg/ml levels. We have also developed a multi-layered normalization process (including positive controls, standard curves, and reference samples) which significantly enhances the performance of the platform and may be used for accurate measurement of protein concentrations in a variety of samples, including serum, plasma, tissue lysates, and other fluids. This platform is also high throughput, allowing hundreds of samples to be assayed daily. We have analyzed several hundred serum samples (both normal and cancer patients) using this system. Our results demonstrate that our technologies provide a powerful approach for delineating candidate biomarkers with potential clinical significance and may be broadly applicable to cancer and other human diseases.

Citation Format: Ruo-Pan Huang, Yingqing Mao, RuoChun Huang, Haw-Han Yen. Quantitative human secretome: a technology to measure the expression levels of thousand of secreted proteins. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3880.
Abstract 3892: Unique pleural vs peritoneal mesothelioma exosomal signature: does mesothelial cell susceptibility to asbestos matter
Authors: Shukla Arti , Munson Phillip , Thompson Joyce K. , Dragon Julie , MacPherson Maximilian B. , Peinado Hector
Published: July 2016

Malignant mesothelioma (MM) is a cancer of mesothelial cells of the pleura or peritoneum caused exclusively by asbestos exposure. It is a deadly cancer which is only diagnosed at terminal stages, possibly due to the long latency period of development and lack of biomarkers for early diagnosis. Both pleural and peritoneal mesothelial cells can develop MM in response to asbestos, however, pleural MM (85%) is more common than peritoneal MM (15%). To get an insight of this difference we first isolated exosomes from pleural (Hmeso, H2373) and peritoneal (ORT) MM cells and performed proteomic analysis. We found that 45 proteins are common and specific to MM cells derived from pleural cavity (Hmeso/H2373) while 86 proteins are specific to ORT cells lines derived from peritoneal cavity. On the other hand, the non-transformed mesothelial cell line, LP9, showed a completely different profile with 269 proteins secreted in exosomes not shared with MM models. This led us to hypothesize that mesothelial cells of two different locations may have different susceptibility to asbestos, accounting for differences observed in pleural and peritoneal MM. To test this hypothesis we exposed human primary pleural and peritoneal mesothelial cells to asbestos for 8 h, and performed massively parallel sequencing on the RNA. Gene expression analysis performed on the RNA-Seq data showed a higher magnitude of responses from pleural mesothelial cells as compared to peritoneal mesothelial cells for the same genes. In addition, at a p<0.05 and 2-fold threshold, unique genes expressed in pleural mesothelium were pro-inflammatory genes known to be involved in MM tumorigenesis. Taken together, results from our study suggest that exosomal cargo from pleural and peritoneal MM cells bear unique signatures which may be due to the different responses of pleural and peritoneal mesothelial cells to asbestos. Future directions involve assessing miRNA cargo from exosomes of pleural and peritoneal MM cells and identifying potential biomarker candidates. This work is supported by grants from NIH (RO1 ES021110), DoD (W81XWH-14-1-0199) and the University of Vermont Cancer Center/Lake Champlain Cancer Research Organization.

Citation Format: Arti Shukla, Phillip Munson, Joyce K. Thompson, Julie Dragon, Maximilian B. MacPherson, Hector Peinado. Unique pleural vs peritoneal mesothelioma exosomal signature: does mesothelial cell susceptibility to asbestos matter. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3892.
Abstract 3896: Novel in vivo pan-Hsp90 family protein inhibitor, Panvotinib-401, showed potent anticancer activities without HSF1 activation
Authors: Kang Byoung Heon
Published: July 2016

Hsp90 family proteins are implicated in tumorigenesis and many inhibitors have been developed as cancer therapeutics. The inhibitors showed potent inhibitory activities against purified Hsp90 family proteins, Grp94 and TRAP1, as well as Hsp90 in vitro. The mitochondrial TRAP1, however, is not inactivated in vivo due to insufficient drug accumulation in the mitochondria. Here, we showed that simultaneous inactivation of Hsp90 family proteins in different compartments can augment anticancer activity of the Hsp90 inhibitors. When simultaneously inhibited, cytoplasmic calcium elevation and activation of calcineurin inhibited HSF1 triggering heat shock responses such as cytoprotective Hsp70 induction. To improve anticancer efficacy of Hsp90 inhibitors, we developed a small molecule (∼380 Da) mitochondrial inner membrane-permeable Hsp90 inhibitor, and named it as Panvotinib-401, briefly Pan-401. Pan-401 showed much improved cancer-selective cytotoxic activity compared with Hsp90 inhibitors or mitochondria-targeted (accumulating) inhibitor, gamitrinib. Collectively, with improved subcellular distribution, cancer-specific cytotoxic activity of Hsp90 inhibitors can be further enhanced.

Citation Format: Byoung Heon Kang. Novel in vivo pan-Hsp90 family protein inhibitor, Panvotinib-401, showed potent anticancer activities without HSF1 activation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3896.
Abstract 3898: Structure-based optimization of small molecule inhibitors of the protein-protein interaction between menin and mixed lineage leukemia (MLL)
Authors: Pollock Jonathan , Borkin Dmitry , Kempinska Katarzyna , Purohit Trupta , Li Xiaoqin , Wen Bo , Zhao Ting , Miao Hongzhi , Shukla Shirish , He Miao , Sun Duxin , Cierpicki Tomasz , Grembecka Jolanta
Published: July 2016

Chromosomal translocations that affect the Mixed Lineage Leukemia (MLL) gene result in acute myeloid and lymphoid leukemias. Fusion of MLL with one of 60 different partner genes generates MLL fusion proteins which lead to enhanced cell proliferation, up-regulation of Hoxa9 and Meis1 genes and block hematopoietic differentiation, ultimately leading to acute leukemia. Importantly, the N-terminal region of MLL is retained in all MLL fusion proteins and represents the interaction site of menin, a critical oncogenic cofactor for MLL fusion proteins. As a result, inhibition of the menin-MLL interaction should abrogate the development and progression of MLL leukemia. Therefore, disruption of the menin-MLL interaction with small molecule inhibitors might represent a therapeutic strategy for patients harboring MLL-rearrangements.

The development of potent small molecule inhibitors of protein-protein interactions with optimized drug-like properties represents a challenging task in the lead optimization process. Here, we employed structure-based design with extensive medicinal chemistry efforts to optimize the thienopyrimidine class of menin-MLL inhibitors represented by the MI-136 compound. Our efforts resulted in development of MI-538, which represents the most potent small molecule inhibitor of the menin-MLL interaction developed to date. MI-538, which binds to menin with a Kd of 6.5 nM demonstrated superior cellular activity in MLL-AF9 transformed leukemic cells with a GI50 = 83 nM. In a mouse xenograft model utilizing MV4:11 human MLL leukemia cells, treatment with MI-538 significantly reduced tumor growth. Overall, systematic exploration of substituents on MI-136 led to the identification of MI-538 with improved activity, selectivity, polarity and pharmacokinetic profile, making it suitable for in vivo studies. Interestingly, we found that simultaneous incorporation of multiple substituents optimized for each site on the cyanoindole ring of MI-136 does not always positively affect the activity or drug-like properties of these compounds. This study demonstrates challenges in optimizing inhibitors of protein-protein interaction for potential therapeutic applications.

Citation Format: Jonathan Pollock, Dmitry Borkin, Katarzyna Kempinska, Trupta Purohit, Xiaoqin Li, Bo Wen, Ting Zhao, Hongzhi Miao, Shirish Shukla, Miao He, Duxin Sun, Tomasz Cierpicki, Jolanta Grembecka. Structure-based optimization of small molecule inhibitors of the protein-protein interaction between menin and mixed lineage leukemia (MLL). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3898.
Abstract 3899: Preclinical development of tetra-branched NT4 peptide theranostics
Authors: Brunetti Jlenia , Depau Lorenzo , Falciani Chiara , Riolo Giulia , Mandarini Elisabetta , Pini Alessandro , Bracci Luisa
Published: July 2016

The tetra-branched peptide NT4 is a potential cancer theranostic, which very selectively binds to human cancer tissues in different malignancies and can efficiently and selectively deliver drugs or liposomes for cancer cell imaging or therapy, in vitro and in vivo. By using NT4 conjugated to methotrexate or 5FdU we obtained significant reduction of tumor growth in xenografted nude mice. Very recently we reported that conjugation of paclitaxel to NT4 leads to increased therapeutic activity of the drug in an orthotopic model of breast cancer in mice and produces tumor regression which is not achieved with unconjugated paclitaxel in identical experimental conditions. We demonstrated that NT4 specifically binds to sulfated glycosaminoglycans and LRP receptors on cancer cells and tissues.

Considering the role of sulfated glycosaminoglycans in cancer cell interaction with the extracellular matrix, we have analyzed the effect of NT4 in cancer cell adhesion and migration on different supports. NT4 inhibits adhesion and migration of different human cancer cell lines, strongly affecting directionality of cell movement.

We have also constructed and validated a novel theranostics nanodevices, by conjugation of NT4 to quantum dots, for selective diagnosis and imaging of different human carcinomas.

Thanks to their high cancer selectivity and versatile chemical conformation, NT4 peptides can be exploited for constructing cancer theranostics, which may also reduce tumor aggressiveness and metastatic potential by inhibiting cancer cell migration.

References:

Falciani, C. et al. Cancer selectivity of tetrabranched neurotensin peptides is generated by simultaneous binding to sulfated glycosaminoglycans and protein receptors. J Med Chem. 2013, 56, 5009-18.

Brunetti, J. et al. Tumor-selective peptide-carrier delivery of Paclitaxel increases in vivo activity of the drug. Scientific Reports. DOI:10.1038/srep17736.

Citation Format: Jlenia Brunetti, Lorenzo Depau, Chiara Falciani, Giulia Riolo, Elisabetta Mandarini, Alessandro Pini, Luisa Bracci. Preclinical development of tetra-branched NT4 peptide theranostics. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3899.
Abstract 3905: Anti-tumor effect of combined irreversible electroporation and liposome-encapsulated NVP-BEZ235
Authors: Li Tian , Wang Lucas , Qiao Yang , Singhana Burapol , Melancon Marites P.
Published: July 2016

Introduction

Irreversible electroporation (IRE) is a technique that uses electrical pulses to cause disruption in the lipid bilayer integrity by creating nanopores. Regions that are close to the probe will have a permanent, irreversible damage to the cells, while farther away regions will subject the cell membranes to re-seal (reversible). This reversible region provides opportunity for enhanced nanoparticle uptake. In this study, we tested the efficacy of combined IRE and nanoparticle-mediated drug delivery in vitro and in vivo in a hepatocellular carcinoma model.

Methods

Liposome-encapsulated NVP-BEZ235 (L-BEZ) was made from hydration-sonication, followed by an optional extrusion step. Particle size was tested via dynamic light scattering. BEZ concentration was quantified by fluorescence. Cytotoxicity of IRE and its drug combinations was tested on Hep3B cells using ECM 830 (BTX Harvard Apparatus) at 1600 V/cm field strength. In vivo efficacy was studied on nude mice bearing Hep3B xenografts with single dose of IRE, or drug alone (L-BEZ or BEZ at 4 mg/kg, intratumoral injection), and its combinations.

Results and Discussion

Liposomes range from 100-500 nm in hydrodynamic volume. Highest loading efficiency achieved was 90% (1.8 mg/mL of BEZ). To test the release of BEZ from the liposome, solution of L-BEZ was treated with IRE and the supernatant was tested against Hep3B cells. The extract from L-BEZ had a significantly higher cytotoxicity (51%) as compared to the empty liposome (20%) with a p-value<0.05. This suggests that IRE can release BEZ from the liposome. The LC50 values of L-BEZ and BEZ was determined to be the same (47 ng/mL). In vitro combination treatment of L-BEZ + IRE showed higher cytotoxicity than the controls (BEZ + IRE, BEZ alone, and IRE alone). In vivo, IRE and its combinations significantly suppressed tumor growth more than non-IRE treated groups. Histology showed a median percent necrosis for L-BEZ + IRE (37%) doubled compared to IRE treatment alone (17%).

Conclusion

The use of minimally invasive IRE in combination with nanoparticle-mediated drug delivery treatment is a promising technique to achieve local disease control in hepatocellular carcinoma.

Citation Format: Tian Li, Lucas Wang, Yang Qiao, Burapol Singhana, Marites P. Melancon. Anti-tumor effect of combined irreversible electroporation and liposome-encapsulated NVP-BEZ235. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3905.
Abstract 3907: Enhanced chemotherapeutic effect with matrix metalloproteinase sensitive liposomes
Authors: Brogaard Rikke Y. , Eliasen Rasmus , Melander Fredrik , Hansen Anders E. , Kjær Andreas , Andresen Thomas Lars
Published: July 2016

Introduction: Drug bioavailability following intra-tumoral accumulation is a major challenge in existing drug delivery systems. A site-specific trigger for obtaining drug release specifically in tumor tissue would overcome this problem and enhance the antitumor effect of the carried drug. Our aim was to develop a new drug delivery platform based on liposomes that are sensitive to matrix metalloproteinases (MMPs).

Methods: In the present work, we have designed a matrix metalloproteinase (MMP)-sensitive liposomal drug delivery system encapsulating oxaliplatin, which by tumor-specific enzymatic dePEGylation allows for controlling the precision of drug release. The dePEGylation is provided by a cholesterol-anchored PEGylated lipopeptide containing a short peptide sequence cleavable by MMP-2 and MMP-9. These MMP-sensitive liposomes are designed to make a charge-reversal transformation upon encountering MMPs in the tumor environment. The surface charge of the liposome turns from slightly negative to positive, resulting in cationic liposomes. In vitro cellular uptake of the liposomes was assessed using flow cytometry and ICP-MS, and further confirmed by confocal microscopy in CT26 and HT1080 cell lines. In vivo studies was evaluated in a syngeneic murine model of MMP-positive CT26 colon cancer.

Results: We successfully formulated MMP-sensitive, oxaliplatin encapsulated liposomes and characterized their zeta potential, size, stability and encapsulation efficiency. The results showed uniformly dispersed particles, and the charge-reversal properties were confirmed as the surface charge changed from slightly negative to distinct positive upon enzymatic cleavage generating cationic liposomes. These MMP-sensitive liposomes were evaluated in vitro and in vivo with a high correlation. In vivo studies showed a prolonged circulation profile with minimal leakage, and similar accumulation in tumors as conventional Stealth liposomes. Efficacy experiment in the same model demonstrated significantly improved antitumor activity relative to free oxaliplatin, and with a superior effect compared to the conventional Stealth liposomes.

Conclusion: With this study, we establish a promising liposomal drug delivery platform for the carriage and release of numerous anticancer drugs into the microenvironment of an MMP-positive tumor.

Citation Format: Rikke Y. Brogaard, Rasmus Eliasen, Fredrik Melander, Anders E. Hansen, Andreas Kjær, Thomas Lars Andresen. Enhanced chemotherapeutic effect with matrix metalloproteinase sensitive liposomes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3907.
Abstract 3908: A multifunctional peptide for targeted imaging, localization of transformed cell and chemotherapy for nasopharyngeal carcinoma and other cancers
Authors: Lin Chin-Tarng , Hsiao Jong-Kai , Liu Hon-Man , Wu Hang-Chung , Yu Alice
Published: July 2016

Introduction: In treating cancer with chemotherapy, the ultimate goal is to deliver sufficient amounts of the drug to tumor cells while minimizing the damage of normal tissue. To achieve this purpose, using a phage-displayed random peptide library to screen the nasopharyngeal carcinoma (NPC)-TW01 cell line, we identified a specific peptide (L-peptide), which binds specifically to NPC tumor cells but not normal cells. To investigate whether this L- peptide can also bind to other cancer cell lines, be used for cancer tissue binding, targeted imaging and chemotherapy, we have performed this experiment. Methods: A fluorescein-labelled L-peptide was used to perform FACScan for 7 cancer cell lines. The L-peptide was linked to dextran coated iron oxide (L-P-Fe3O4) nanoparticles and a biotin labeled modified L-peptide (biotin-mL-P) were also used to localize L-peptide targeted protein (X-protein) in NPC and breast cancer surgical specimens. This conjugate was also injected intravenously to SCID mice bearing NPC and breast cancer xenografts for magnetic resonance imaging (MRI) analysis. Furthermore, L-peptide-linked liposomal doxorubicin (L-P-Lipo-Dox) was used to observe the efficacy of peptide-targeted chemotherapy in both cancers. Results and Conclusions: FACscan revealed specific binding of L-peptide to NPC, breast cancer, lung cancer, and neuroblastoma. The X-protein in both cancer specimens localized by biotin-mL-P was seen in many NPC and breast cancer tumor cells with some unstained tumor cells, but not or very weak in normal tissue; the X-protein in formalin-fixed paraffin-embedded section could be localized by L-P-Fe3O4 histochemistry in both cancer specimens. MRI analysis of both cancer xenograft bearing mice after injection of L-P-Fe3O4 revealed significant change in the MR signal intensity. A high efficacy of L-P-Lipo-Dox treatment for both cancers with minimal adverse effect was obtained. This multifunctional L-peptide seems able to be used for localization of its binding protein in each cancer cell type, molecular MRI analysis and targeted chemotherapy with minimal adverse effect in different cancer types.

Citation Format: Chin-Tarng Lin, Jong-Kai Hsiao, Hon-Man Liu, Hang-Chung Wu, Alice Yu. A multifunctional peptide for targeted imaging, localization of transformed cell and chemotherapy for nasopharyngeal carcinoma and other cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3908.
Abstract 3932: Values of single nucleotide polymorphisms identified from genome-wide association studies on risk prediction, risk reclassification, and population-based screening of breast-cancer
Authors: Huang Yubei , Song Fengju , Chen Kexin
Published: July 2016

Background: With an increasing number of single nucleotide polymorphisms (SNPs) being identified from genome-wide association studies (GWAS), it's important to examine whether these SNPs have values in public health. And it's also unclear how to select the targeted SNPs valuable for population-based screening from numerous GWAS-identified SNPs.

Methods: Based on the fraction of the genetic risk explained by a SNP, the area under the receiver operating characteristic curve (AUC), integrated discrimination improvement, and net reclassification improvement were used to select targeted SNPs and to evaluate the values of genetic risk score (GRS) with targeted SNPs on risk prediction, risk reclassification, and effects of population-based screening among 2 million simulated Chinese women aged 35-69 years.

Results: A total of 11 targeted SNPs from 23 GWAS-identified SNPs would be valuable for population-based screening of breast cancer among Chinese women. After incorporating GRS with targeted SNPs into traditional risk predication based on established risk factors (ERF) of breast cancer, the AUC significantly improved from 65.5% to 67.5% (P<0.001). Additional 25.5% (P<0.001) population would be correctly reclassified into cases or controls. If screening targeted at top 25% high-risk women, screening-detected cancer would significantly increase 4.2% (49.2% vs 45.0%, P<0.001) for screening strategy included GRS and ERF in risk assessment before screening compared with that only included ERF in risk assessment.

Conclusion: GRS based on targeted GWAS-identified SNPs could significantly improve the risk prediction, risk reclassification, and the effects of population-based screening. Further real-world studies are needed to validate this method and these results.

Citation Format: Yubei Huang, Fengju Song, Kexin Chen. Values of single nucleotide polymorphisms identified from genome-wide association studies on risk prediction, risk reclassification, and population-based screening of breast-cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3932.
Abstract 3934: Analysis of PKC-ζ protein and mRNA levels in normal and malignant breast tissue
Authors: Apostolatos Christopher , Smalley Tracess , Duncan Mildred A.
Published: July 2016

It is estimated that in 2015 breast cancer will be the second leading cause of cancer death in women. For this purpose, biochemical markers for breast cancer have been investigated, to assist in early detection and more accurate diagnoses. In breast cancer tissue, the atypical isozyme of protein kinase C zeta, PKC-ζ, has been a topic in research; It has been shown that an overexpression of this protein may be indicative of developing carcinogenesis and contributes to proliferation through the NF kappa B pathway, which is a stress-regulated switch for cell survival. In this investigation, the expression of PKC-ζ was analyzed in normal and malignant female human breast tissue samples by Western blot, immunoprecipitation and PCR. In the preliminary results, the malignant breast tissue samples illustrated a significant expression of PKC ζ when compared to the expression of PKC-ζ in normal breast tissue samples. The same tissues were also processed for total RNA isolation which was followed by cDNA synthesis and Real Time PCR. The level of PKC-ζ mRNA was tested and no overexpression was observed in either malignant or normal breast tissue samples. While protein studies may suggest that PKC-ζ could be considered a biomarker for breast cancer, the same cannot be said about mRNA levels. The overexpression of PKC-ζ protein level and the normal PKC-ζ mRNA level are indicators of possible miRNA activity that may regulate translation in malignant tissues but not the normal.

Citation Format: Christopher Apostolatos, Tracess Smalley, Mildred A. Duncan. Analysis of PKC-ζ protein and mRNA levels in normal and malignant breast tissue. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3934.
Abstract 3944: Combining tumor glypican-3 expression and CD16 Expression on NK cells from peripheral blood to identify patients responding to codrituzumab/GC33/RO5137382
Authors: Chen Gong , Chen Ya-Chi , Reis Bernhard , Belousov Anton , Jukofsky Lori , Rossin Christine , Muehlig Axel , Xu Chao , Essioux Laurent , Ohtomo Toshihiko , Di Laurenzio Laura , Puig Oscar , Lee Ray
Published: July 2016

Background & Aims: Codrituzumab (GC33 or RO5137382), a monoclonal antibody targeting an oncofetal protein glypican-3 (GPC3) expressed on the cell surface of hepatocellular carcinoma (HCC), induces antibody-dependent cellular cytotoxicity (ADCC) and inhibits tumor growth in preclinical studies. Based on this mechanism of codrituzumab, two biomarkers GPC3 and CD16 of the NK cells, which are the effector cells of ADCC, were investigated to correlate with clinical efficacy of codrituzumab in patients with advanced HCC.

Methods: Data from a phase II clinical trial of codrituzumab were used for this analysis. GPC3 expression was determined by immunohistochemistry (IHC) analysis of tumor biopsies, and CD16 expression on NK cells were quantified by a flow cytometry analysis of peripheral blood mononuclear cells. Overall survival of patients with high exposure of codrituzumab or placebo (n = 113) was used to compare different patient subgroups stratified by high or low expression of GPC3 and CD16.

Results: In individual-biomarker analyses, longer survival after codrituzumab treatment correlates with either high expression level of GPC3 in tumors (n = 97, HR = 0.39, 95%CI = 0.21 - 0.70, p = 0.00081) or a relatively high level of CD16 expressed on NK cells (n = 80, HR = 0.44, 95%CI = 0.23 - 0.83, p = 0.0056) at baseline. Furthermore, joint analyses of the two biomarkers reveal that both high levels of GPC3 and CD16 are required for patients to benefit from codrituzumab treatment (n = 67, HR = 0.29, 95%CI = 0.13 - 0.62, p = 0.00074), and lack of either one of them leads to a loss of codrituzumab therapeutic effect.

Conclusions: The retrospective analysis supports the mechanism of ADCC, in which the combination of high GPC3 expression in tumors and high CD16 expression in NK cells from peripheral blood is associated with prolonged overall survival given treatment of codrituzumab. This result supports the usage of both GPC3 and CD16 as potential biomarkers to select HCC patients for codrituzumab treatment.

Citation Format: Gong Chen, Ya-Chi Chen, Bernhard Reis, Anton Belousov, Lori Jukofsky, Christine Rossin, Axel Muehlig, Chao Xu, Laurent Essioux, Toshihiko Ohtomo, Laura Di Laurenzio, Oscar Puig, Ray Lee. Combining tumor glypican-3 expression and CD16 Expression on NK cells from peripheral blood to identify patients responding to codrituzumab/GC33/RO5137382. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3944.
Abstract 395: Patient stratification and drug combination strategy based on drug response and genomic information from PDX clinical trials (PCTs)
Authors: Jiang Jingjing , Yu Tengfei , Yan Ying , Du Wei , Tan Tingting , Yang Xuqin , Gu Jiali , Ye Xin K. , Gu Zhenyu
Published: July 2016

Cancer is a heterogeneous disease with various molecular lesions and drug response profiles within the same tumor type. Patient stratification in clinical trials based on molecular features has contributed to recent success of several targeted cancer drugs on molecular aberrations such as BCR-ABL translocation in CML, Her2 amplification in breast and gastric cancers, EGFR mutations and ALK fusions in lung cancer and BRAF mutation in melanoma. However, in most cancers, the molecular features which can be used for patient stratification are not as simple as a single genetic aberration. Multiple drug resistance mechanisms caused by various mutations in cancer signaling pathways can also increase the uncertainty of clinical outcomes.

To increase the chance of success in human clinical studies, patient-derived xenograft (PDX) clinical trials (PCTs) have increasingly been used for predictive biomarker validation, resistance mechanism investigation and combination therapy selection. PDX tumor models have been demonstrated to have high correlations with human patients in tumor pathology, molecular characteristics and drug responses. Large scale PCTs have also shown consistency in results when compared to related human clinical trials.

At GenenDesign, we have established over 1000 PDX tumor models and more than 100 resistance models against various cancer drugs. Many of these PDX models have been characterized at RNA/Exome sequence, gene expression, gene copy number and hot-spot mutation levels. We carried out our PDX clinical trials by testing multiple approved drugs and clinical drug candidates such as targeted inhibitors against FGFRs, c-Met/ALK, HER2, EGFR, cell cycle regulators, Ras/Raf pathway, PI3K/Akt pathway, as well as chemotherapy drugs in biomarker-driven multi-drug multi-arm expanded PDX clinical trials. So far, we have accumulated more than 3000 efficacy data sets and associated PD samples.

Analysis of drug response and associated genomic information from PDX clinical trials yielded rich information for predictive biomarker identification and validation. At the same time, many potential resistance mechanisms were also revealed. These information can make human clinical trial better prepared, more efficient and focused. More importantly, testing of a targeted drug with multiple chemotherapies in the same models can also provide guidance on future combination selection strategy.

Citation Format: Jingjing Jiang, Tengfei Yu, Ying Yan, Wei Du, Tingting Tan, Xuqin Yang, Jiali Gu, Xin K. Ye, Zhenyu Gu. Patient stratification and drug combination strategy based on drug response and genomic information from PDX clinical trials (PCTs). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 395.
Abstract 3950: Optimization of a method for FACS analysis of checkpoint receptor expression on tumor infiltrating lymphocytes in clinical tumor samples
Authors: Zhang Norman , Zhang Qiyao , Liu Yan , Yu Juntao , Wang Jingjing , Xu Jie , Ji Qunsheng , Wilcoxen Keith , Graves Jonathan
Published: July 2016

While immune checkpoint therapies against CTLA-4, PD-1 and PD-L1 have achieved promising clinical outcome recently, the dynamic therapeutic responses spanning individual patients and different cancer types warrant exploration and validation of additional therapeutic strategies. Since the expression of immune checkpoints in tumor infiltrating lymphocytes (TIL) has been of interest to the field, we attempted to analyze the expression of several checkpoint receptors on TILs in clinical tumor samples. Unexpectedly, a number of markers were subject to time-dependent degradation during the tissue disaggregation process. This degradation could be due to the proteolytic activity of a proteinase, which has been used commonly in various tissue disaggregation protocols. Analysis of the protein sequences verified the presence of the proteinase recognition sequences in the affected receptors. Using PHA-stimulated hPBMCs, we found and demonstrated that a specific protease was responsible for either partial or total degradation of these markers. To overcome this issue, we developed a protease-free tissue disaggregation method, which showed no adverse effect on protease affected cell membrane receptors, thus allowing for more reliable flow cytometric analysis of various TIL subpopulations. With this method, we analyzed these checkpoint receptors positivity of both CD4+ and CD8+ TILs and revealed a high degree of heterogeneity of these immune checkpoints in a number of tumor types, including lung, breast and gastric cancers. Analysis of the magnitude of lymphocyte infiltration to the tumor and expression of prominent immune checkpoints on CD4+ or CD8+ TILs will help understand the subtle interaction between tumor and the immune system in the tumor microenvironment and may provide guidance for clinical trials of novel combinatorial immuno-therapeutic strategies.

Citation Format: Norman Zhang, Qiyao Zhang, Yan Liu, Juntao Yu, Jingjing Wang, Jie Xu, Qunsheng Ji, Keith Wilcoxen, Jonathan Graves. Optimization of a method for FACS analysis of checkpoint receptor expression on tumor infiltrating lymphocytes in clinical tumor samples. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3950.
Abstract 3954: Prospective evaluation of circulating tumor cells (CTCs) in head and neck cancer patients receiving definitive radiotherapy with a nanotechnology based system
Authors: Caster Joseph M. , Eblan Michael J. , Wang Kyle , Myung Ja Hye , Chera Bhishamjit , Hong Seungpyo , Wang Andrew Z.
Published: July 2016

Background: Circulating tumor cells (CTCs) are an important biomarker in cancer. There has been substantial interest in utilizing CTCs to enable personalized treatment. However, there is limited data on CTCs in patients with head and neck cancers (HNC) where there is growing data that curative treatment needs to be risk-based. The purpose of this prospective, correlative study is to investigate the clinical significance of CTCs, as measured by a highly sensitive CTC capture technology UiChip, in HNC patients undergoing definitive treatment.

Methods: HNC patients (M0) undergoing definitive treatment (radiation+/- chemotherapy) were enrolled. Peripheral blood was collected prior to starting RT, at the first week, mid-point, final week of treatment, and every 4 to 12 weeks after completion. Quantification of CTCs was conducted using UiChip. The primary endpoint was change in CTCs pre- and post- RT.

Results: 35 patients are included in this analysis (median age 58, 34% non-smokers, 69% HPV/p16 positive, and 17% node (-)) and 90% received chemotherapy. CTCs were detected in all patients pre-RT (100%). There was no association between pre-RT CTC level and tumor or nodal stage. Median CTCs significantly decreased from 71 CTCs/mL (range, 7-849) before RT to 32 (2-209) at completion of RT and 27 (2-78) at 4 to 12 weeks post-treatment. Ten patients had persistent disease and/or distant metastasis at a median follow-up of 10.6 months (range, 3.7-17). CTCs declined throughout treatment in patients with complete clinical and/or radiographic responses, in contrast to an elevation in CTCs at mid or post-RT in the 7 patients with pathologic residual disease/distant failures.

Conclusions: We demonstrated that the novel UiChip can capture CTCs in a diverse population of head and neck cancer patients. Our pilot data suggest that individual patient CTC changes during and after treatment may be a predictive biomarker for radiotherapy response and clinical outcomes. Table: Median CTCs/mL Peripheral Blood (Median% of Pre-Treatment Baseline Count)Pre-RTMid-RTPost-RTNumber (%) of CTCs in Complete Responders to RT (N = 18)67 IQR: 39-11737 (38%) IQR: 22-79 (18-94%)21 (18%) IQR: 7-40 (13-42%)Number (%) of CTCs in Partial Responders to RT (N = 7)120 IQR: 36-30366 (123%) IQR: 20-134 (30-172%)51 (45%) IQR: 23-54 (17-63%)

Citation Format: Joseph M. Caster, Michael J. Eblan, Kyle Wang, Ja Hye Myung, Bhishamjit Chera, Seungpyo Hong, Andrew Z. Wang. Prospective evaluation of circulating tumor cells (CTCs) in head and neck cancer patients receiving definitive radiotherapy with a nanotechnology based system. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3954.
Abstract 3978: Lipid tethering to enable real-time imaging of breast tumor cell cytoskeletal dynamics and rapid drug testing in free-floating metastatic microenvironments
Authors: Chakrabarti Kristi Riti , Andorko James , Whipple Rebecca , Zhang Peipei , Sooklal Elisabeth , Jewell Christopher , Martin Stuart
Published: July 2016

Free-floating tumor cells located in the peripheral circulation of cancer patients, known as circulating tumor cells (CTCs), play an important role in cancer and have become key targets for studying metastasis. The presence of CTCs in the bloodstream or lymphatics correlates with decreased cancer patient survival and provides a minimally invasive method to study disease progression and treatment response. Understanding the molecular characteristics and functional properties of CTCs have been impeded by the challenges of imaging CTCs under conditions that model the free-floating microenvironments of the circulation. Upon extracellular-matrix (ECM) detachment, breast tumor cells form tubulin-based protrusions known as microtentacles (McTNs) that play a role in the aggregation and re-attachment of tumor cells to increase their metastatic efficiency. In this study, we have designed a strategy to spatially immobilize ECM-detached tumor cells within a microfluidic device while maintaining their free-floating character. We use cytophobic polyelectrolyte multilayers deposited on microfluidic substrates to prevent tumor cell adhesion and the addition of lipid moieties to tether tumor cells to these surfaces through interactions with the cell membrane. This approach enables high-resolution time-lapse microscopy of McTNs on viable free-floating tumor cells and real-time analysis of dynamic cellular features. In addition, tethering makes it now possible to rapidly measure drug responses of tumor cells in free-floating microenvironments to select effective therapies and avoid drugs that could inadvertently increase metastasis. The ability to image tumor cells in the absence of ECM attachment can immensely enhance our understanding of CTCs under conditions that better recapitulate the free-floating microenvironments that tumor cells encounter during metastasis.

Citation Format: Kristi Riti Chakrabarti, James Andorko, Rebecca Whipple, Peipei Zhang, Elisabeth Sooklal, Christopher Jewell, Stuart Martin. Lipid tethering to enable real-time imaging of breast tumor cell cytoskeletal dynamics and rapid drug testing in free-floating metastatic microenvironments. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3978.
Abstract 3982: Prediction and selection of cancer drug treatments using personalized tumor models or models with matching genomic profiles
Authors: Jiang Jingjing , Yan Ying , Luo Zhongguang , He Jia , Yu Tengfei , Du Wei , Yang Xuqin , Gu Jiali , Ye Xin K. , Zhuang Guanglei , Liu Jie , Gu Zhenyu
Published: July 2016

The main purpose of precision medicine is to find the right drug for the right patient at the right time. Current progress in cancer drug target identification and development of new targeted drugs has presented many successful examples. However, for majority of patients, finding treatments based on the unique mutations in their own tumor cells are still hard. New approaches for precision medicine are in great needs and thus being investigated.

A mouse “avatar”, also known as Patient-Derived Xenograft (PDX) model, is a personalized cancer model derived from a patient tumor sample and used to test different drugs for the patient. “GIFTS” (or Genomic Information Fitting based Therapeutics Selection) is a method with which a patient's cancer genomic information is compared to PDX model genomic profiles to find the best genomic fit and related therapeutic options in GenenDesign Drug Response Database and Genomic Information database.

So far we have successfully derived more than 1000 PDX models from patient tumor tissues of various cancer types including lung, gastric, liver, esophageal, colorectal, pancreatic and many other cancers. In some cases, both patients and their avatars were tested with same cancer drugs including targeted drugs and chemotherapies. By comparing drug responses in mouse avatars with patient clinical results, we found high correlations between them in both sensitivity and unresponsiveness.

We have also developed a bioinformatics algorithm to analyze PDX model genomic and drug response information. Both drug sensitivity and resistance biomarkers have been used in matching cancer patient genomic profiles to those of PDX models in GenenDesign database as a part of our GIFTS method. Our preliminary results show that there are high degree of similarities in drug response profiles between patients and their matched PDX models.

Avatar and GIFTS methods can provide predictions of therapeutic effectiveness on both targeted drugs and chemotherapies. Avatar is a drug screening based method, while GIFTS is a genomic profile matching based method. Both methods are especially useful for patients, whose tumors could not be treated with known targeted drugs.

Citation Format: Jingjing Jiang, Ying Yan, Zhongguang Luo, Jia He, Tengfei Yu, Wei Du, Xuqin Yang, Jiali Gu, Xin K. Ye, Guanglei Zhuang, Jie Liu, Zhenyu Gu. Prediction and selection of cancer drug treatments using personalized tumor models or models with matching genomic profiles. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3982.
Abstract 3986: Diagnostic significance of cathepsin L and cathepsin B expression in human gallbladder cancer - A pilot study
Authors: MEHRA SIDDHARTH , KUMAR MANISH , PANWAR RAJESH , DASH NIHAR RANJAN , SINGH RATNAKAR , YADAV RAJNI , GUPTA SIDDHARTHA DATTA , SAHNI PEUSH , CHAUHAN SHYAM S.
Published: July 2016

Background: Gallbladder cancer (GBC) is the most common biliary tract malignancy worldwide. Cancer of gallbladder usually refers to as an Indian disease as it is more common in Indian subcontinent especially in north and central India. Incidence rate of GBC in women in Delhi is as high as 21.5 per 100,000 population. It is an aggressive malignancy, which spreads locally to liver, adjacent organs and lymph nodes as well as to distant organs. Cathepsin L (CTSL) and B (CTSB) are lysosomal cysteine proteases implicated in protein turnover and tissue remodeling. Deregulated expression of these proteases has been associated with invasion and metastasis in several solid tumors.

Objective: To assess the expression and clinical significance of CTSL and CTSB in human gallbladder cancer.

Methods: We assayed the activities of CTSL and CTSB via spectroflurometrically in whole tissue lysates of gallbladder cancer patients (N = 23) and normal gall bladder tissue samples with gallstones (N = 31) / without gallstones (N = 34), which served as controls. Activities of these proteases were correlated with several clinic pathological parameters in GBC patients. CTSL and CTSB mRNA levels were determined in tissues samples by Real time PCR. Furthermore serum levels of these proteases were also assayed using sandwich based ELISA in GBC patients and controls.

Results: Significant increase in the enzymatic activities of CTSL+B, CTSL and CTSB was observed in gallbladder cancer tissue samples in comparison to normal gall bladder both with and without gallstones (p < 0.001). ROC analysis confirmed the diagnostic relevance of CTSL and CTSB analysis with area under the curve 0.791 and 0.840 respectively for gallbladder cancer patients with respect to controls. Over expression of CTSB was also significantly associated with involvement of lymph node (p = 0.05) in gall bladder carcinoma. Secretory levels of CTSL and CTSB in serum were significantly elevated in GBC patients (p <0.01). Increased expression of these cysteine proteases were also confirmed at the mRNA level by Real time PCR.

Conclusion: Our study for the first time demonstrated a significant increase in the levels of cathepsin L and B in GBC patients as compared to controls. Elevated levels of these proteases may serve as potential biomarkers for human gall bladder cancer.

Citation Format: SIDDHARTH MEHRA, MANISH KUMAR, RAJESH PANWAR, NIHAR RANJAN DASH, RATNAKAR SINGH, RAJNI YADAV, SIDDHARTHA DATTA GUPTA, PEUSH SAHNI, SHYAM S. CHAUHAN. Diagnostic significance of cathepsin L and cathepsin B expression in human gallbladder cancer - A pilot study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3986.
Abstract 3993: Th1 cytokines regulate apoptotic cell death and HER family RTK expression in murine and human breast cancer lines
Authors: Namjoshi Prachi M. , Showalter Lori , Czerniecki Brian , Koski Gary K.
Published: July 2016

A recent neoadjuvant vaccine trial to treat early breast cancer demonstrated powerful induction of Th1 immunity against the HER-2, complete pathologic responses in over 18% of subjects, and for many subjects, evidence of down-regulated HER-2 expression on residual disease. To explain these observations, we investigated the action of archetypical Th1 cytokines (TNF-α + IFN-γ) on both murine and human breast cancer cell lines that varied in the surface expression of HER-family receptor tyrosine kinases. We found that most tumor cell lines were sensitive to dual Th1 cytokines as evidenced by lower metabolic activity (alamar blue assay), lower proliferation, and enhanced apoptosis (AnnexinV/PI staining and TUNEL assay) as well as a reversible inhibition of surface expression of HER proteins. Apoptotic cell death was accompanied by demonstrated increases in activated caspase-3. Furthermore, the pharmacologic caspase-3 activator, procaspase-activating compound (PAC-1), mimicked both the killing effects and HER-2 suppressive activities of Th1 cytokines, while the caspase 3/7 inhibitor ((5-[(S)-(+)-2-(Methoxymethyl)pyrrolidino]sulfonylisatin), prevented cytokine-induced HER-2 downregulation. These studies therefore demonstrated that many of the in vivo effects of vaccination (apparent tumor cell death and loss of HER-2 expression) could be replicated in vitro using only the principle Th1 cytokines. These findings are consistent with the notion that IFN-γ and TNF-α work in concert to mediate some of the biological effects of therapeutic Th1-polarizing vaccination through the induction of a caspase 3-dependent cellular death mechanism.

Citation Format: Prachi M. Namjoshi, Lori Showalter, Brian Czerniecki, Gary K. Koski. Th1 cytokines regulate apoptotic cell death and HER family RTK expression in murine and human breast cancer lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3993.