bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2766: Automation of linear range determination: Enzymatic progress curve applications
Authors: May Earl William , Urul Daniel , Cornell-Kennon Susan , Lu Zhibing , Schaefer Erik , Hoare Sam , Laliberté France , Vong Quay , Payette Paul , Marois Jean 
Published: April 2023

Abstract
To characterize modulators of enzymatic activity, a continuous assay format will quantify with confidence the initial reaction velocity far better than an end point assay. When the progress curve is linear from the start and remains linear throughout the experiment, a single end point reading can approximate the reaction rate, but a kinetic assay will provide the rate more accurately. However, if there is a delay in the onset of the reaction, or there are other changes in reaction rate over time, then a single end point reading is insufficient. Thus, important information about the reaction can be missed or mischaracterized. While visual inspection is the most common way to determine the range of the progress curve from which to extract rate information, this method is not practical when thousands of progress curves are generated in an experimental day. We have developed an automated protocol to streamline and optimize the process. The heuristic algorithm analyzes the progress curve to identify different regions for characterization (e.g. lag, primary rate, secondary rate, final plateau) and then extract rates of interest for each of these regions. These rates are subsequently used to measure the specific activity of the enzyme under various assay conditions, and to characterize the modulation of that activity: simple dose-response testing, mode of inhibition analysis, time-dependent inhibition determination, reversibility testing, determination of residence time for reversible inhibitors, and kinact/KI analysis for irreversible inhibitors. We describe the algorithm and several applications as they apply to protein kinases using the PhosphoSens assay platform from AssayQuant within the Analyze module of the Scigilian data analysis package.
Citation Format: Earl William May, Daniel Urul, Susan Cornell-Kennon, Zhibing Lu, Erik Schaefer, Sam Hoare, France Laliberté, Quay Vong, Paul Payette, Jean Marois. Automation of linear range determination: Enzymatic progress curve applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2766.
Abstract 2786: Targeting oncogene and inflammation for cancer therapy
Authors: Wang Wei , Datta Sayantap , Velu Sadanandan E. , Luo Ming , Zhang Ruiwen 
Published: April 2023

Abstract
Accumulating evidence has demonstrated that the overexpression of and the loss of tumor suppressor genes contribute to the aggressiveness, poor prognosis, and poor response to treatment associated with advanced cancer. Overexpression and activation of the MDM2 and NFAT1 oncogenes frequently occur in human cancer and is associated with invasive and high grade/late-stage tumors, metastasis, and recurrence, and are negative prognostic factors for cancer patients. Dual targeting MDM2 and NFAT1 represents a novel molecular target for the treatment of cancer. However, most of the current MDM2 inhibitors have been designed to block the binding between MDM2 and p53, and have limited efficacy against tumors with mutant or deficient p53. Considering that high incidence of TP53 mutations and MDM2 overexpression in tumors, these MDM2 SMIs are expected to have low or no efficacy against those types of cancer. Therefore, there is an urgent need to identify a novel, p53-independent strategy to inhibit MDM2, in order to affirm the therapeutic value of targeting MDM2. In addition, as an important inflammatory factor, there are no specific NFAT1 inhibitors developed for cancer therapy. By employing crystallization screening, NMR-based screening, and high-throughput virtual and cell-based screening assays, we discovered a class of makaluvamine analogs with the desired property. One of these analogs, MA242, was identified as a potent and selective MDM2 and NFAT1 inhibitor. Its in vitro and in vivo anti-breast cancer activities and underlying mechanisms of action were evaluated in breast cancer cell lines with various p53 backgrounds. Our results demonstrated that MA242 directly bound to MDM2 and NFAT1 protein with high affinity and induced its protein degradation. As a result of this binding, MA242 decreased cell proliferation, and induced apoptosis and G2/M phase arrest in breast cancer cell lines, regardless of the p53 status. Furthermore, MA242 inhibited the tumor growth and metastasis in orthotopic and patient-derived xenograft (PDX) models, without any host toxicity. Single-cell sequencing results provided the molecular and cellular crosstalk between cancer and immune system, and identified the predictive biomarkers of response after the treatment. In conclusion, MA242 is a potent and selective MDM2 and NFAT1 dual inhibitor with distinct mechanisms of action compared to the existing inhibitors. Our data suggest that the dual targeting oncogene and inflammation is a promising strategy for the treatment of human cancer. (Supported by NIH R01CA214019 and a UH DDI Seed Grant.)
Citation Format: Wei Wang, Sayantap Datta, Sadanandan E. Velu, Ming Luo, Ruiwen Zhang. Targeting oncogene and inflammation for cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2786.
Abstract 2796: Tumor pharmacokinetics, pharmacodynamics and efficacy analysis of WEE1 inhibitor, zn-c3 in patient-derived xenograft models of glioblastoma
Authors: Mehta Shwetal , Tien An-Chi , Patel Sonam , Margaryan Tigran , Shivanna Anilkumar Thaghalli , Elena Leonel , Molloy Jennifer , Knight William , Hon Ruiheng , Elliott Mackenna , Stavnichuk Mariya , Opachich Zorana , Chang Yu-Wei , Montgomery Chelsea , Himes Sarah , Hopkins Barbara , Tovmasyan Artak , Sanai Nader 
Published: April 2023

Abstract
WEE1 kinase regulates G2/M checkpoint transition via phosphorylation of CDK1 and prevents entry into mitosis. WEE1 inhibition increases genomic instability due to G2/M checkpoint inactivation and can result in mitotic catastrophe as monotherapy or when combined with DNA damaging agents. In this study, we evaluated the efficacy as well as pharmacokinetic and pharmacodynamic properties of ZN-c3, a best-in-class WEE1 inhibitor, in orthotopic patient-derived xenograft (PDX) models of GBM. Mice with intracranial tumors were randomized to 80 mg/kg ZN-c3 PO QD x 2 weeks vs. placebo. ZN-c3 levels in plasma and contrast-enhancing tumor tissue were measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). Unbound fractions were determined by equilibrium dialysis. Immunohistochemistry was performed to assess levels of pCDK1, phospho-Histone H3 (pHH3), and cleaved caspase 3 (CC3). The median unbound concentrations of ZN-c3 were 315 nmol/L and 15 nmol/kg in plasma and tumor tissue, respectively - higher than the biochemical IC50 of ZN-c3 for inhibition of WEE1 (3.8nM). An increase in pHH3(+) cells (0.40% vs. 6.57%, p=0.03) indicated drug-induced mitotic entry upon ZN-c3 treatment. ZN-c3 was potently cytotoxic across multiple patient-derived xenograft (PDX)-derived cell lines (IC50- 150-10000nM). Time and dose-escalation experiments exhibited target engagement at concentrations lower than IC50 values within one hour of treatment. Basal levels of WEE1 and pCDK1 were correlated with cell viability in response to ZN-c3 across multiple PDX cell lines. Overall, ZN-c3 is well tolerated, achieves pharmacologically-relevant unbound concentrations in GBM PDX models, and is associated with significant checkpoint modulation. Preclinical evaluation of ZN-c3 in combination with radiation therapy and DNA damage response pathway inhibitors is currently underway, which will support future combinatorial strategy for glioblastoma treatment.
Citation Format: Shwetal Mehta, An-Chi Tien, Sonam Patel, Tigran Margaryan, Anilkumar Thaghalli Shivanna, Leonel Elena, Jennifer Molloy, William Knight, Ruiheng Hon, Mackenna Elliott, Mariya Stavnichuk, Zorana Opachich, Yu-Wei Chang, Chelsea Montgomery, Sarah Himes, Barbara Hopkins, Artak Tovmasyan, Nader Sanai. Tumor pharmacokinetics, pharmacodynamics and efficacy analysis of WEE1 inhibitor, zn-c3 in patient-derived xenograft models of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2796.
Abstract 2804: 3D printing of perfusable alveolar model for efficacy evaluation of polyisoprenylated cysteinyl amide inhibitors on lung cancer cells
Authors: Romero Diana Isabel , Ren Bing , Leon Christopher , Offringa Ite A. , Lamango Nazarius S. , Huang Yong 
Published: April 2023

Abstract
Lung cancer is the leading cause of cancer death in the United States, and Black Americans are being disproportionality affected. Lung adenocarcinoma (LUAD) is the most common histological lung cancer subtype among all lung cancer patients, including black patients. There is still no available physiologically relevant three-dimensional (3D) in vitro model that can be used to study the treatment efficacy on LUAD cells, in particular, those from Black patients. In this study, we aim to develop a 3D alveolar model with a perfusable epithelialized channel using 3D biofabrication technologies. We will apply the model to test their efficacies of polyisoprenylated cysteinyl amide inhibitors (PCAIs) and other drugs on LUAD. First, a perfusable channel is constructed within a gelatin composite extracellular matrix (ECM) using a sacrificial material-assisted 3D embedded printing technology. Next, the channel is seeded with KRAS-mutated NCI-H23 LUAD (derived from a Black patient) or hAEC-C7 (untransformed, immortalized human alveolar epithelial cells) respectively, and exposed to drugs to test the anti-cancer efficacy. In particular, PCAIs, which are potential therapeutics targeting activated KRAS, are investigated for their cytotoxicity on H23 LUAD and hAEC-C7 cells for comparison. Compared to two-dimensional (2D) models, the proposed 3D alveolar model can offer a more physiologically relevant 3D cell-to-ECM environment to test LUAD therapeutics. The use of H23 cells and, in the future, other LUAD cells from Black patients ensures that any developed drugs constitute valid treatment options for black patients with RAS-mutated pathways, thus minimizing health disparities.
Citation Format: Diana Isabel Romero, Bing Ren, Christopher Leon, Ite A. Offringa, Nazarius S. Lamango, Yong Huang. 3D printing of perfusable alveolar model for efficacy evaluation of polyisoprenylated cysteinyl amide inhibitors on lung cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2804.
Abstract 2833: Exploring the impact of IL-17 pathway modification on radiation dermatitis and radiation therapy efficacy
Authors: Pirtle Megan E. , Kost Yana , Deutsch Alana , Shinoda Kosaku , McLellan Beth N. 
Published: April 2023

Abstract
Here we explore the role of Interleukin-17 (IL-17) in the promotion of radiation dermatitis (RD) and radiation therapy (RT) efficacy. RD is a common and potentially treatment-limiting skin toxicity that occurs following RT. When the treated tumor is close to the skin, as in head and neck or breast cancer, RD is the most common acute adverse event which negatively impacts patients’ quality of life; however, evidence-based treatments are limited. We performed RNA sequencing of RT-treated skin in both human skin and a murine model, which each showed increased expression of IL-17 Receptor type C (IL-17RC). To further explore the role of the IL-17 pathway in the development of RD, both genetic knockout of IL-17RC and pharmacologic blocking of IL-17A in mouse models were studied, and both were found to have attenuated histopathological signs of RD, including inflammation, acanthosis, and fibrosis when compared to controls. After establishing the importance of the IL-17 pathway on RD, we explored the impact of such IL-17 pathway modification on the anti-tumorigenic efficacy of RT, an important step when considering a future IL-17 neutralizing therapy for RD. A Trans-PyMT murine model of triple-negative breast cancer was used to evaluate the impact of IL-17A neutralization on RT efficacy. There was no difference found in the efficacy of RT in mice treated with IL-17A neutralizing antibody as compared with saline control. Overall, we present a clinically relevant murine model which advances our understanding of the immunopathogenesis of RD and suggests further exploration of targeted therapies for RD prophylaxis is warranted. Additionally, we show that modifying the IL-17 pathway is effective in mitigating RD phenotype with no impact on RT efficacy.
Citation Format: Megan E. Pirtle, Yana Kost, Alana Deutsch, Kosaku Shinoda, Beth N. McLellan. Exploring the impact of IL-17 pathway modification on radiation dermatitis and radiation therapy efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2833.
Abstract 2845: PpIX synthesis increases PDT resistant cells due to the modulating effect of EGCG on the heme synthesis pathway
Authors: León Daniela , Gutiérrez Álvaro , Weber Helga , Silva Ramón , Reyes María Elena , Viscarra Tamara , Buchegger Kurt , Ili Carmen , Brebi Priscilla 
Published: April 2023

Abstract
Introduction: Photodynamic therapy (PDT) is a treatment for non melanoma skin cancer. PDT kills cells through reactive oxygen species (ROS), produced by interaction between cellular O2 and an excited photosensitizer (PS) by a specific light. One of the most used PS in dermatology is Protoporphyrin IX (PpIX) and its precursor methyl aminolevulinate (MAL). MAL enters into cells and is converted to PpIX by enzymes of the heme group synthesis pathway. The insertion of ferrous iron into PpIX forms a heme molecule, which has no PS action. A previous study shows that efficacy of PDT, in resistant cells, can be improved by addition of epigallocatechin gallate (EGCG), a green tea polyphenol. Therefore, the aim of this work is to evaluate the rol of EGCG on PPIX synthesis.
Material and methods: It was used a A-431 cell line from skin squamous cell carcinoma. The EC50 of EGCG was determined by MTT assay after 24 hours. Clonogenic assay was carried out using EGCG 10, 20 and 40 μM, for 4 and 24 hours of incubation. To evaluate PpIX and ROS, A-431 cells were exposed to EGCG (10, 20, 40 μM) with or without MAL 2 mM. After 4 hours, some plates were irradiated with red light to generate ROS. PpIX and ROS were detected by flow cytometry. Gene expression of enzymes of heme synthesis was analyzed by RT-qPCR. Intracellular iron was measured using a commercial kit. ﻿
Results: EC50 determined a concentration of 50 μM of EGCG at 24 hours. EGCG concentrations from 10 μM reduced cell clonogenic capacity, even with only 4 hours of incubation. A-431 cells show PDT resistance with MAL 2 mM and a fluence of 4 J/cm2. The addition of EGCG to MAL-PDT showed a decrement of cell viability with an increase of PpIX and ROS. Besides, incubation with EGCG for 4 hours, triggered an increment of HMBS enzyme and decrease of FECH. In addition, EGCG reduced intracellular iron concentrations.
Conclusion: These findings show that EGCG has a PDT enhancing effect, decreasing intracellular iron levels and modulating gene expression of heme synthesis enzymes, which are directly related with high levels of PpIX production and ROS levels, causing the death of resistant cells.Acknowledgements: Postdoctoral FONDECYT Nº 3210618, IDeA I+D FONDEF ID21I10027
Citation Format: Daniela León, Álvaro Gutiérrez, Helga Weber, Ramón Silva, María Elena Reyes, Tamara Viscarra, Kurt Buchegger, Carmen Ili, Priscilla Brebi. PpIX synthesis increases PDT resistant cells due to the modulating effect of EGCG on the heme synthesis pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2845.
Abstract 2875: Evidence of immunosuppression in patients with breast ductal carcinoma in situ
Authors: Taghinezhad-S Sedigheh , Mohseni Amir Hossein , Beatty Pamela L. , Emens Leisha A. , Finn Olivera J. 
Published: April 2023

Abstract
Introduction: Ductal carcinoma in situ (DCIS) is a noninvasive precursor to invasive breast cancer. Although not all DCISlesions progress to cancer, most DCIS are treated as early breast cancer, given the uncertainty of which lesions will progress. The current increase in the incidence of DCIS has sparked interest in the immune contexture of the DCIS lesions as well as the immune status of DCIS patients, as a means to predict which DCIS lesions will progress to cancer. Studies done so far in invasive breast cancer showed that the immune system can mediate both tumor-promoting influence (via T regulatory (Treg) cells, M2 macrophages, myeloid-derived suppressor cells (MDSCs)), as well as tumor-suppressing influences (via CD8+ and CD4+ effector T cells, natural killer cells (NK), M1 macrophages, tissue-resident memory T (Trm) cells, B cell and dendritic (DCs) cells). Similar studies are still lacking in DCIS. Our aim was to start closing the gap in our understanding of the DCIS immune microenvironment, which will pave the way for novel preventative and therapeutic strategy.
Methods: We analyzed by flow cytometry peripheral blood mononuclear cells (PBMCs) from newly diagnosed DCIS patients for the presence of various immune cell types expressing specific phenotypic markers of immune effectors or immune suppressors and compared them with PBMC from age-matched healthy individuals.
Results: We found a statistically significant increase in the frequency of FoxP3+ regulatory T cells in DCIS PBMC compared with healthy controls. There was also a significant increase in Tim-3+CD8+ T effector memory cells (P= 0.035) and Tim-3+ CD8+ NKT cells (P = 0.033) in the PBMCs of patients with DCIS compared with healthy controls. T cells expressing other exhaustion markers, such as PD-1+ and LAG-3+ were present at very low levels in both groups.
Conclusion: Our results suggest that some of the many immunosuppressive mechanisms seen in patients with invasive breast cancer, such as regulatory T cells, are already present in pre-cancer DCIS. Evidence of the beginning of immune suppression is also reflected in the increased frequency of CD4+ and CD8+ T cells expressing the exhaustion marker Tim-3.
Citation Format: Sedigheh Taghinezhad-S, Amir Hossein Mohseni, Pamela L. Beatty, Leisha A. Emens, Olivera J. Finn. Evidence of immunosuppression in patients with breast ductal carcinoma in situ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2875.
Abstract 3068: CREBBP/EP300 disruption promotes tumor progression and confers synthetic lethality in anaplastic thyroid cancers
Authors: Haase Jacob , Gebhard Talia A. , Liu Qi , Bernabé Sara G. , Hao Jingzhu , Unegbu Chisom , Bikas Athanasios , Qi Jun , Landa Iñigo 
Published: April 2023

Abstract
Anaplastic thyroid cancers (ATC) are fast-growing, undifferentiated tumors and almost invariably fatal, primarily due to the lack of effective therapeutic options. The recent approval of dabrafenib plus trametinib for the treatment of BRAFV600E-mutant ATCs improved the prognosis of a subset of patients, but ineligibility and acquired resistance still limit their use. Overall, ATC patients remain in great need of tailored therapeutic options. We previously showed that loss-of-function alterations targeting histone acetyltransferase (HAT) genes, namely CREBBP and EP300, occur in 15-20% of ATCs, but only in &lt;1% of their well-differentiated counterparts. What remains unknown are the specific mechanisms by which HAT disruptions perturb chromatin architecture, impact gene homeostasis, and unleash cellular processes in these aggressive tumors. We are assessing the HAT-mediated thyroid cancer progression. CREBBP/EP300 knockouts enhanced thyroid cancer cell proliferation in vitro and induced thyroid gland growth in a thyroid-specific mouse model of HAT loss. We are leveraging these animals to characterize the in vivo effects of Crebbp/Ep300 knockout, alone or in combination with BrafV600E, in thyroid cancer phenotypes and epigenetic reconfiguration. In addition, we are exploiting the molecular consequences of HAT loss to explore tailored treatments. CRISPR/Cas9 screens identified a mutual CREBBP/EP300-dependency of HAT-mutant human cancer cell models. Our experiments in thyroid cancer cells employing CREBBP/EP300-targeting proteolysis-targeting chimera (PROTAC) compounds specifically degraded these proteins and decreased histone acetylation. Our findings prove the oncogenicity of HAT loss in thyroid cancer progression and support exploring synthetic lethality dependencies in CREBBP/EP300-mutant ATCs. In summary, we provide pre-clinical basis to inform genomics-driven and mechanism-oriented decisions for the clinical management of patients with HAT-altered ATC.
Citation Format: Jacob Haase, Talia A. Gebhard, Qi Liu, Sara G. Bernabé, Jingzhu Hao, Chisom Unegbu, Athanasios Bikas, Jun Qi, Iñigo Landa. CREBBP/EP300 disruption promotes tumor progression and confers synthetic lethality in anaplastic thyroid cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3068.