bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2162: ActivSignal technology for multiplex analysis of signaling protein activation profiles
Authors: Alexandrov Ilya , Mackenzie Malcolm , Brandina Irina 
Published: August 2019

Abstract
The IPAD assay technology from ActivSignal monitors activity of 26 major signaling pathways in multiplex by examining phosphorylation status or expression of 70 protein targets from canonical signaling pathways. The technology uses antibody pairs for each protein target to ensure high specificity, and these pairs are carefully selected and tested. Only if both antibodies in a pair bind to their specific target molecule will the detection take place during the analysis phase - avoiding false hits. The ActivSignal IPAD platform can be used to profile cell culture samples, tissues and different blood fractions. Human and mouse samples can be analyzed. The IPAD assay requires only a very small amount of your precious sample. Applications include:
- Direct analysis of protein activity of the signaling cascades and not subsequent downstream events.
- Testing physiological effects of compounds of interest.
– Influence on specific protein targets, as well as the signaling pathways as a whole.
– Secondary and/or off-target effects.
- Similarly, analysis of signaling pathways activation status in response to different stimuli, such as: CRISPR, shRNA, hormones, stresses and other treatments.
- Medium-throughput analysis (96-well format).
- Tissue sample profiling from patients and animals (murine).
- Profiling of tumor biopsies.
Our collaborators analyzed the mechanism of action of JG-98, an Hsp70 inhibitor that exhibits anti-cancer activities that affect both cancer cells and tumor associated macrophages. Using gene expression and ActivSignal protein profiling assay, the signaling pathways influenced by JG-98 were identified, which predicted potential drug combinations that can be used with JG-98 for further preclinical development. ActivSignal is a breakthrough, low-cost tool for the analysis of signaling pathway activation profiles. It enables researchers to rapidly and efficiently gain insight into function of their favorite drug molecules, genes etc.
Citation Format: Ilya Alexandrov, Malcolm Mackenzie, Irina Brandina. ActivSignal technology for multiplex analysis of signaling protein activation profiles [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 2162.
Abstract 2169: Doxorubicin conjugation to reovirus enhances tumor cell-directed oncolysis
Authors: Berry Jameson T. , Mainou Bernardo A. 
Published: August 2019

Abstract
Breast cancer remains the most diagnosed type of cancer and second leading cause of cancer-related deaths in women in the United States. Triple-negative breast cancer (TNBC) constitutes 10-20% of breast cancer cases. TNBCs are characterized by lack of estrogen (ER) and progesterone (PR) receptors and growth factor receptor HER2/Neu on the cell surface. Therapies that target ER, PR, or HER2 are thus ineffective against TNBC. TNBC is primarily treated with cytotoxic chemotherapy or radiation therapy, which can cause significant damage to healthy cells and tissue. Therefore, there is need for therapies that selectively kill TNBC cells with reduced off-tumor effects. The concept of oncolytic virotherapy has existed since the early 1900s. However, oncolytic viruses have limited efficacy as single agents. Mammalian orthoreovirus (reovirus) is a non-enveloped, segmented double-stranded RNA virus with tropism for transformed cells. Reovirus is in Phase I - III clinical trials to test its oncolytic efficacy against a variety of cancers. Success has been limited and few trials have tested reovirus against breast cancer. In a high-throughput screen of small molecule inhibitors, we identified the topoisomerase II inhibitor doxorubicin hydrochloride (dox) as an enhancer of reovirus infectivity in the TNBC cell line MDA-MB-231. To better control dox delivery and enhance reovirus oncolytic potential, we chemically conjugated dox to reovirus virions (reo-dox). While reo-dox attachment to MDA-MB-231 cells is slightly impaired, viral replication is largely unaffected. Infection of MDA-MB-231 cells with reo-dox resulted in increased cytopathicity and faster induction of cell death than virus alone. MDA-MB-231 cells infected with reo-dox, but not virus alone, induced DNA double-strand breaks and activation of DNA damage response. Together, our findings show that reo-dox exhibits superior toxicity in TNBC cells than virus alone. Future studies will define the mechanism of enhanced cytopathicity of reo-dox and oncolytic efficacy of dox-conjugated reovirus in vivo. Delivery of small molecule inhibitors via conjugation to reovirus particles may provide an effective new method to directly target and kill cancer cells while minimizing toxicity to healthy cells and tissues.
Citation Format: Jameson T. Berry, Bernardo A. Mainou. Doxorubicin conjugation to reovirus enhances tumor cell-directed oncolysis [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 2169.
Abstract 2173: Developing a high-throughput, high content screening technology to identify new anticancer drug treatments
Authors: Milcarek Andrew T. , Daus Kevin , Alpaugh Mary L. 
Published: August 2019

Abstract
The high attrition rate of new chemotherapeutic drugs represents the single most important challenge in anticancer drug development. With only one drug out of 10, 000 test compounds gaining FDA-approval, attrition is often attributed to failure of in vitro drug efficacy to translate to clinical success. The overall drug discovery and development pipeline can be dramatically improved with new, innovative assays of high pharmacological relevance that can shorten the discovery/preclinical phase and ensure rapid translation to clinic. We report the development of a three-dimensional phenotypic drug screening platform based on spheroidsMARY-X. The tight, compact spheroidsMARY-X form spontaneously due to innate molecular determinants with proliferative cells on the outer periphery and a dormant tumor cell core. These features mimic the intratumoral biological complexities of tumor tissue and thus provide a relevant in vitro model for drug discovery with a high degree of predictive value. Significantly, this model allows analysis of drug targeting of dormant tumor cells, the difficult to treat resistant or highly refractory cell population. Drug response can be reliably assessed using brightfield microscopy and can be quantified by measuring spheroid circularity (i.e., dissolution index). Evaluation of soluble factor concentration is performed using molecular rotors through restricted intramolecular rotation upon protein binding with a subsequent quantum yield. Overall, we present a 3-dimensional anticancer assay that provides a rapid, efficient, relevant platform for the evaluation of drug response and allows identification of small molecule hits in both proliferative and more importantly, dormant tumor cells, the major cause of relapse and recurrent metastatic disease.
Citation Format: Andrew T. Milcarek, Kevin Daus, Mary L. Alpaugh. Developing a high-throughput, high content screening technology to identify new anticancer drug treatments [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 2173.
Abstract 2175: Synthetic antigen receptor mesenchymal stem cells (SAR-MSCs) targeting perlecan for drug delivery to ovarian cancer
Authors: Nethi Susheel Kumar , Sehgal Dristhi , Cheng Shen , Panyam Jayanth , Prabha Swayam 
Published: August 2019

Abstract
Mesenchymal stem cells (MSCs) can be engineered with polymeric nanoparticles for tumor-targeted delivery of small molecule drugs. Such nano-engineered MSCs have demonstrated exciting anticancer activity in multiple ovarian cancer models. Despite significantly improved delivery of chemotherapeutics to tumor tissues, non-specific accumulation of MSCs in clearance organs remains a concern. Paralleling the concept of CAR-T cells, we advance here a strategy for synthetic modification of MSCs with antibodies targeting specific antigens overexpressed on cancer cells. Our approach consists of stably incorporating recombinant protein G (PG) on the surface of MSCs, followed by binding of a full-length IgG to the PG handle. Because protein G binds to the Fc region of IgG, antigen-binding affinity of the antibody is conserved. We have previously shown the overexpression of perlecan (HSPG2) on ovarian cancer cells and its correlation to poor patient survival. In the current study, we investigated the incorporation of anti-perlecan antibody on the surface of nano-engineered MSCs. The anti-perlecan IgG antibody was first derivatized with palmitic acid (PA), which was then used to insert PG on MSCs cell membrane. We characterized the PAPG derivative by LC/Q-TOF/MS. Flow cytometry and confocal microscopy were used to confirm the incorporation of fluorescently labeled PAPG handle on MSC surface. Similarly, the binding of fluorescently labeled anti-perlecan IgG to PAPG-functionalized MSCs was confirmed by flow cytometry. We are currently investigating the anticancer efficacy of paclitaxel-loaded, anti-perlecan SAR-MSCs using in vitro and mouse models of ovarian cancer.
Citation Format: Susheel Kumar Nethi, Dristhi Sehgal, Shen Cheng, Jayanth Panyam, Swayam Prabha. Synthetic antigen receptor mesenchymal stem cells (SAR-MSCs) targeting perlecan for drug delivery to ovarian 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 2175.
Abstract 2177: Streptavidin-pHast: A readily conjugatable, pH-sensitive dye to screen for internalization
Authors: Shramm Patrick A. , Ancheta Leonardo 
Published: August 2019

Abstract
Quick and efficient screening of targeting agents that internalize effectively is vital for determining their suitability as potential therapeutics. Some of the most recent successes in the treatment of cancers have been from antibodies to cell surface proteins that are responsible for tumor cell proliferation. Examples are Cetuximab (target: EGFR) approved for colorectal cancer, and Trastuzumab (target: HER2) for breast cancer. These antibodies have more than one effect on the cancer cell, but one of the most important is that, upon binding to the cell surface antigen, the complex is internalized. As such, the down-regulated cell surface protein no longer plays a role in cancer cell division. Here we describe a method for determining internalization of cell surface molecules by targeting agents using a pH-dependent fluorescent reporter cross-linked to streptavidin. Streptavidin is a tetrameric protein (molecular weight 53 kDa in its recombinant form), with each subunit able to bind a single biotin molecule. The bond between streptavidin and biotin is rapid and essentially non-reversible, unaffected by most extremes of pH, organic solvents, and denaturing reagents. It is the strongest known noncovalent biological interaction (Ka = 1015 M-1) between protein and ligand. A variety of molecules, including lectins, proteins, and antibodies, can be biotinylated and reacted with streptavidin-labeled probes or other detection reagents for use in biological assays. The fluorescence from this reporter increases intensity as the pH of its surroundings becomes more acidic, as demonstrated when exposed to the environment inside a cell (thereby providing evidence of internalization). Here we describe methods that can be used to explore candidates as cancer therapeutics in a quick, reliable and reproducible manner.
Citation Format: Patrick A. Shramm, Leonardo Ancheta. Streptavidin-pHast: A readily conjugatable, pH-sensitive dye to screen for internalization [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 2177.
Abstract 2182: Preclincial in vitro and in vivo combination therapies for mutant IDH1R132 tumors with BAY 1436032
Authors: Kaulfuss Stefan , Zaman Julia , Chaturvedi Anuhar , Hess-Stumpp Holger , Höde Jennifer , Biber Ricarda , Borowicz Renan , Heuser Michael , Deimling Andreas von , Pusch Stefan 
Published: August 2019

Abstract
Background: Isocitrate dehydrogenase (IDH) mutations are drivers of different tumors. The IDH1R132 mutation is frequently found in glioma (80%), chondrosarcoma (CS, 45%), intrahepatic cholangiocarcinoma (ICC, 20%), acute myeloid leukemia (AML, 7%), and with low incidences in other solid tumors. Although clinical therapies of mutant IDH inhibition were recently approved for the treatment of AML, few patients benefit from a single therapy with a mutant IDH inhibitor and most of them relapse. Here we report preclinical in vitro and in vivo data on combination therapies of the mIDH1R132 inhibitor BAY 1436032 with standard of care chemotherapies.
Methods: in vitro: The sarcoma cell line HT1080 (IDH1R132C), together with two IDH1 wildtype sarcoma cell lines, the SW1353 (IDH2R172S) and SW872 (IDHwt) cell lines, were treated with BAY 1436032 alone and in combination with different standard of care drugs. in vivo: The patient derived xenograft (PDX) ICC model LIXFC2084 (IDH1R132L), the melanoma PDX model MEXF1341 (IDH1R132C) (MEXF1341), the PDX AML model 13PB020 (IDH1R132C) and a sarcoma cell line derived xenograft model HT1080 (IDH1R132C) were included. All models were treated with BAY 1436032 alone and in combination with different standard of care (SOC) drugs.
Results: The in vitro treatment with BAY 1436032 unexpectedly increased growth in HT1080 (IDH1R132C) cells with no effect to the control cell lines SW1353 and SW872. In contrast, in vivo, combination of BAY1436032 with everolimus had additive anti-tumor efficacy and was synergistic with cisplatin or the MEK inhibitor refametinib in the treatment of HT1080 xenografts. The combination of BAY 1436032 and refametinib was also additive in the treatment of a melanoma xenograft with IDH1R132C mutation. Another additive efficacy of BAY 1436032 and gemcitabine or cisplatin could be observed in an ICC xenograft model and further in the combination with everolimus in the same model. The combination of BAY 1436032 together with azacitidine in a PDX AML model was also synergistic.
Conclusion: These results highlight the combinatorial potential of the mIDH1R132 inhibitor BAY 1436032 with standard of care therapy and the importance of microenvironmental factors, which has to be taken into account when dealing with mutant IDH tumors.
Citation Format: Stefan Kaulfuss, Julia Zaman, Anuhar Chaturvedi, Holger Hess-Stumpp, Jennifer Höde, Ricarda Biber, Renan Borowicz, Michael Heuser, Andreas von Deimling, Stefan Pusch. Preclincial in vitro and in vivo combination therapies for mutant IDH1R132 tumors with BAY 1436032 [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 2182.
Abstract 2183: Protective autophagy elicited by RAF®MEK®ERK inhibition suggests a treatment strategy for pancreatic cancer
Authors: Kinsey Conan G. , Camolotto Soledad A. , Boespflug Amelie M. , Guillen Katrin P. , Foth Mona , Truong Amanda , Schuman Sophia S. , Shea Jill E. , Seipp Michael T. , Yap Jeffrey T. , Burrell Lance D. , Lum David H. , Whisenant Jonathan R. , Gilcrease G. Weldon , Cavalieri Courtney C. , Rehbein Kaitrin M. , Cutler Stephanie L. , Affolter Kajsa E. , Welm Alana L. , Welm Bryan E. , Scaife Courtney L. , Snyder Eric L. , McMahon Martin 
Published: August 2019

Abstract
Pancreatic ductal adenocarcinoma was responsible for ~43,000 deaths in the USA in 2017 and is the epitome of a recalcitrant cancer driven by an, as yet, pharmacologically intractable oncoprotein, KRAS. Although the clinical picture remains grim, the mechanisms by which key alterations in tumor suppressors and proto-oncogenes contribute to PDA have been dissected. Downstream of KRAS, the RAF→MEK→ERK signaling pathway plays a central role in pancreatic carcinogenesis. However, to date, pharmacological inhibition of this pathway has provided no clinical benefit to PDA patients. Here we show that inhibition of KRAS→RAF→MEK→ERK signaling in PDA cell lines elicits autophagy, a process of cellular recycling that protects pancreatic cancer cells from the potentially cytotoxic effects of KRAS pathway inhibition. Furthermore, combined inhibition of MEK1/2 plus autophagy displays synergistic anti-proliferative effects against PDA cell lines in vitro, and promotes regression of xenografted patient-derived PDA tumors in mice. Finally, treatment of a PDA patient with the combination of trametinib plus hydroxychloroquine resulted in a partial, but nonetheless striking disease response. These data suggest that this combination therapy may represent a novel strategy to target RAS-driven cancers such as PDA.
Citation Format: Conan G. Kinsey, Soledad A. Camolotto, Amelie M. Boespflug, Katrin P. Guillen, Mona Foth, Amanda Truong, Sophia S. Schuman, Jill E. Shea, Michael T. Seipp, Jeffrey T. Yap, Lance D. Burrell, David H. Lum, Jonathan R. Whisenant, G. Weldon Gilcrease, Courtney C. Cavalieri, Kaitrin M. Rehbein, Stephanie L. Cutler, Kajsa E. Affolter, Alana L. Welm, Bryan E. Welm, Courtney L. Scaife, Eric L. Snyder, Martin McMahon. Protective autophagy elicited by RAF®MEK®ERK inhibition suggests a treatment strategy for pancreatic 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 2183.
Abstract 2185: Efficacy of Rac/Cdc42 inhibitors in multiple cancer models
Authors: Calderon Jean F. Ruiz , Castillo-Pichardo Linette , Lopez-Lopez Irmaris , Dharmawardhane Suranganie 
Published: August 2019

Abstract
The RhoGTPases Rac and Cdc42 are excellent therapeutic targets for inhibiting metastasis because these homologous proteins regulate numerous cellular functions that promote tumorigenesis, angiogenesis, invasion and therefore, metastasis. Also, Rac1, Cdc42, and their effector p21-activated kinase (PAK) are overexpressed in multiple cancer types, including breast, gastric, and pancreatic, and are associated with higher mortality rates. Furthermore, Rac and Cdc42 are activated through the signaling of oncogenic cell surface receptors such as EGFR and HER2, therefore hyperactivating their signaling pathways. Our laboratory has developed two small molecules targeting Rac and Cdc42, Ehop-016 and MBQ-167, that inhibit Rac1 and Cdc42 with IC50s in the low micro- and nano-molar concentrations, respectively (Montalvo-Ortiz et al. 2012, J Biol Chem; Humphries-Bickley et al. 2017, Mol Cancer Therap), making them the most potent Rac/Cdc42 inhibitors developed to date. We hypothesize that targeting Rac1, 3, and Cdc42 is a rational strategy to inhibit aggressive TNBC, pancreatic and gastric cancer. To test this hypothesis, we treated the TNBC cell lines MDA-MB-231, MDA-MB-468 and 4T1 (murine breast cancer), gastric cancer cells NCI-N87, and pancreatic cancer cells MIA-PaCa-2 with 4uM and 500nM of Ehop-016 and MBQ-167 respectively, and measured cell viability. Both TNBC cell lines and the pancreatic cancer cell line exhibited a drastic reduction in cell viability (&gt60%) after 48hrs of treatment Ehop-016 or MBQ-167. Additionally, experimental and spontaneous metastasis assays were performed with TNBC and gastric cancer xenografts in immunocompromised and immunocompetent mice. Treatment with 1 mg/kg BW MBQ-167 3X week resulted in reduced tumor growth by 60-90% in TNBC and gastric cancer models, and metastasis inhibition by ~90% in the MDA-MB-231 model. In the syngeneic experimental metastasis study with 4T1, both Ehop-016 and MBQ-167 reduced lung metastasis in comparison to the vehicle treated group. Moreover, these compounds were not toxic to both immunocompromised and immunocompetent mice, as observed by no significant changes in weight and liver enzymes. Therefore, both Ehop-016 and MBQ-167 are promising targeted therapeutics against cancers that overexpress Rac, Cdc42, and PAK.
Citation Format: Jean F. Ruiz Calderon, Linette Castillo-Pichardo, Irmaris Lopez-Lopez, Suranganie Dharmawardhane. Efficacy of Rac/Cdc42 inhibitors in multiple cancer models [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 2185.
Abstract 2187: Tryptophan Metabolism Plays a Central Role in Immunosuppression
Authors: Geeganage Sandaruwan , Sams Lillian , Henry James , Dorsey Frank , Roth Kenneth , Nikolayev Alexander , Benhadji Karim , Gilmour Raymond , Cerezo Ana , Peregrina Sandra , Hoyo Gloria Martínez-del , Campos-Olivas Ramón , Funes Juan Manuel , Diezma Laura , Velasco-Miguel Susana 
Published: August 2019

Abstract
Tryptophan metabolism plays a central role in immunosuppression through the local depletion of tryptophan with the concomitant production and accumulation of kynurenine, both of which are immunosuppressive. Indoleamine 2,3-dioxygenase 1 (IDO1), a heme-dependent enzyme, catalyzes the initial and rate-limiting step of the kynurenine pathway. Tumor cells selectively upregulate IDO1 as an immune-evasion mechanism either through intrinsic expression of IDO1, or in response to IFN-γ, a cytokine secreted by immune cells during an active immune response.
In addition to the immuno-suppressive role of IDO1 and kynurenine, we investigated their additional roles in tumor cells. Here we describe an NMR-based readout assay for IDO1 activity, in which we are able to trace the extra- and intracellular destination of the single tryptophan carbon groups. This assay allowed us to detect contribution of the tryptophan catabolism to purine synthesis in tumor cells, and suggested further roles of tryptophan catabolism in tumors, apart from kynurenine production, that contribute to the modulation of tumor growth. We detected the incorporation of labeled tryptophan carbon units in intracellular purine pools. Our highly potent, orally available, and CNS penetrant small molecule inhibitor of IDO1 (LY3381916) inhibits this incorporation of tryptophan-derived one carbon units into purines in vitro and in vivo. This activity leads to pre-clinical efficacy and may be an added advantage of inhibiting IDO1 in cancer.
Citation Format: Sandaruwan Geeganage, Lillian Sams, James Henry, Frank Dorsey, Kenneth Roth, Alexander Nikolayev, Karim Benhadji, Raymond Gilmour, Ana Cerezo, Sandra Peregrina, Gloria Martínez-del Hoyo, Ramón Campos-Olivas, Juan Manuel Funes, Laura Diezma, Susana Velasco-Miguel. Tryptophan Metabolism Plays a Central Role in Immunosuppression [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 2187.