Cancer Cell Division Research Articles

Prognostic significance and expression pattern of glucose related genes in breast cancer: A comprehensive computational biology approach

Breast cancer (BC) is the most common type of malignancy globally and the main reason why women die from tumours. The Warburg effect, a characteristic of tumor, describes how most solid tumour cells acclimatize to their diverse surroundings by relying heavily on aerobic glycolysis for production of energy. In addition to producing key metabolic intermediates that are crucial for the production of macromolecules, which enable cancer cell division, invasiveness, and drug resistance, the transformed energy metabolism also supplies tumor cells with ATP for cellular energy. Here, we evaluated the expression profile, prognostic significance, and clinical relevance of glucose-related genes in BC using a bioinformatic approach. To clarify the significance of glucose-related genes in the development of breast tumours, we also performed a functional enrichment investigation of deregulated genes using the STRING and KEGG portal. The study depicted that of the 61 genes examined, 8 genes had a fold change </=± 1.5, that is, ADH1C, ADH4, ALDH1A3, ALDOC, FBP1, PCK1, PFKFB1, PFKFB3. Among the highly deregulated genes, ADH1C showed a fold change of −6.669. These deregulated genes were associated with poor prognosis. The study signifies that glucose related genes are highly dysregulated in breast cancer. Deregulation of glucose related genes is linked with a poor prognosis in BC individuals. Thus, targeting glucose related genes will provide an effective treatment approach for BC individuals.

Eph signal inhibition potentiates the growth-inhibitory effects of PLK1 inhibition toward cancer cells

Anti-mitotic drugs are clinically used as anti-cancer treatments. Polo-like kinase 1 (PLK1) is a promising target against cancer cell division due to its importance in the whole process of mitosis, and thus PLK1-targeting agents have been developed in the last few decades. Clinical trial studies show that several PLK1 inhibitors are generally well-tolerated. However, the response rates are limited; therefore, it is needed to improve the efficacy of those drugs. Here, we show that NVP-BHG712, an erythropoietin-producing human hepatocellular (Eph) signaling inhibitor, potentiates the growth-inhibitory effects of the PLK1 inhibitors BI2536 and BI6727 in cancer cells. This combination treatment strongly suppresses cancer spheroid formation. Moreover, the combination drastically arrests cells at mitosis by continuous activation of the spindle assembly checkpoint (SAC), thereby inducing apoptosis. SAC activation caused by the combination of NVP-BHG712 and BI2536 is due to the inhibition of centrosome maturation and separation. Although the inactivation level of the PLK1 kinase is comparable between BI2536 treatment alone and combination treatment, the combination treatment strongly inactivates MAPK signaling in mitosis. Since inhibition of MAPK signaling potentiates the efficacy of BI2536 treatment, inactivation of PLK1 kinase and MAPK signaling contributes to the strong inhibition of centrosome separation. These results suggest that Eph signal inhibition potentiates the effect of PLK1 inhibition, leading to strong mitotic arrest via SAC activation and the subsequent reduction of cancer cell survival. The combination of PLK1 inhibition and Eph signal inhibition will provide a new effective strategy for targeting cancer cell division.

Synthesis, crystal structure, Hirshfeld surface analysis and DFT calculations of novel clusianone analogues as anticancer agents that downregulate β-tubulin and Cdk1/Cyclin B1

Clusianone is a natural product that has been shown to possess anticancer activity, however, studies on its potential anticancer activity remain limited and few. In this study, we further investigated the anticancer potential of clusianone by initially isolating clusianone from Garcinia parvifolia leaves and then using it to synthesise a total of 8 analogues, of which 5 are novel and being reported for the first time. Clusianone and its analogues’ reactivity and chemistry were thoroughly investigated, while the crystal structure of methylated clusianone, compound 1, was obtained and discussed. The crystal structure for 1 was used to conduct Hirshfeld surface analysis whereby it was found that H···H interactions contributed the most to the crystal packing. Moreover, DFT calculations showed that the calculated theoretical parameters for the optimised structure of 1 were in good agreement with the experimental x-ray diffraction measurements. The analogues that were obtained in good yields were further screened against A549 (lung) and HK1 (nasopharyngeal) cancer cells via the MTT assay. Results showed that introduction of an imine (C=N) group to clusianone resulted in bioactive compounds, with the methylamine derivative (compound 4) demonstrating the highest cytotoxic activity followed by clusianone. Further studies on clusianone and 4 showed that they caused apoptosis-related morphological changes in the cancer cells. Furthermore, the compounds were found to inhibit cancer cell division via decreasing the protein expression levels of β-tubulin and Cdk1/Cyclin B1 with 4 demonstrating a more pronounced effect against the proteins than clusianone. DFT calculations on 4 suggested that its high bioactivity could be attributed to its high chemical reactivity due to its lower HOMO-LUMO gap and lower chemical hardness. This study demonstrates the potential of 4 as an effective natural product derived anticancer agent.

Saponins Derived from the Korean Endemic Plant Heloniopsis koreana Inhibit Diffuse-type Gastric Cancer Cells.

Diffuse-type gastric cancer (GC) is a type of stomach cancer that occurs in small clusters of cells that are widely spread. It does not typically manifest with symptoms until the advanced stages and often goes undetected in routine imaging tests. In addition, there is no specific targeted therapy for diffuse-type GC and it has a high mortality risk. Hence, it is worthwhile to discover molecules against this cancer. In this study, the extract of Heloniopsis koreana, which is endemic to Korea, exhibited cytotoxicity against two diffuse-type GC cell lines, MKN1 and SNU668. This led to the isolation of 10 compounds, including a new cinnamic acid glycoside. Of the compounds, saponin Th (4) and SNF 11 (5) showed potent activities with IC50 values of 3.66 and 3.85 μM, respectively, in MKN1 cells, and 1.8 and 1.98 μM, respectively, in SNU668 cells. These compounds prevented cancer cell division, invasion, and colony formation in both cell lines. In addition, these compounds induced cancer cell death through conventional cell death pathways, showing an increase in ADP-ribose polymerase, caspase 3, and BAX and a decrease in BCL2.

Efficacy and Safety of Polatuzumab Vedotin in Follicular Lymphoma: A Systematic Review of Clinical Trials

Introduction: Polatuzumab vedotin (Pola) is a humanized anti-CD79b monoclonal antibody linked with antimitotic agent monomethyl auristatin E. On internalization, the linker is cleaved, and antimitotic activity inhibits the cell division and growth of cancer cells. Pola is tested in multiple combinations in relapsed/refractory (RR) follicular lymphoma (FL) in recent years. In this systematic study, we will assess the efficacy and safety of Pola based combinations in RR FL. Methods: A literature search was performed on PubMed with keywords, “Polatuzumab vendotin” and “lymphoma” from the inception of data till 07/01/2023. PRISMA guidelines were followed while conducting this systematic review. We reviewed 160 articles and included 1 randomized clinical trial (RCT, N=41), and 4 non-randomized clinical trials (nRCT, N=163) based on inclusion criteria. Data was extracted for complete response (CR), partial response (PR), overall response (ORR), progression free survival (PFS), overall survival (OS) and ≥grade 3 side effects. Results: In 5 trials (N=204), 13 patients with Pola + atezolizumab (Ate) + obinutuzumab (Obi), 49 patients with Pola + Obi + venetoclax (Ven), 39 patients with Pola + Obi, 62 with Pola + Obi + lenalidomide (Len), and 20 patients with Pola + rituximab (Rit). In an RCT by Morschhauser et al. (N=41), Pola + Rit was compared with pinatuzumab vedotin (Pina) + Rit. ORR, CR, PR, and mPFS was 70%, 45%, 25%, and 15.3 months, respectively with Pola + Rit, as compared to 62%, 5%, 57%, and 12.7 months, respectively, with Pina + Rit. In a trial by Philips et al. (N=39), ORR, CR, PR, and mPFS were 67%, 36%, 31%, and 11.5 months, respectively. In 3 nRCTs (N=124), three drug regimens were used with Pola and Obi as backbone. ORR, CR, PR were 50%-83%, 20%-61%, and 20%-39%. Table 1. ≥ Grade 3 hematological and non-hematological side effects are given in table 1. Conclusion: Pola was well tolerated by most of the patients with FL alone or in combination with Obi, Rit, Len, Ven. Pola had relatively better response and survival rates as compared to Pina in combination with Rit. In early phase clinical trials, three drug regimens of Pola and Obi with Ven, and Len were effective in the treatment of heavily pretreated follicular lymphoma. In an early phase trial, Pola + Obi + Ate had limited efficacy in FL with safety concerns. More randomized double blind multicenter phase III clinical trials are needed to confirm these results.

EXTH-27. ASSESING THE EFFECT OF ELECTRIC FIELD THERAPY ON PATIENT DERIVED GLIOMA STEM CELL LINES AND GLIOMA INFILTRATING CEREBRAL ORGANOIDS

Abstract Glioblastoma (GBM) is the most aggressive primary brain tumour in adults. Electric field therapy (EFT) is a new cancer treatment applied after safe maximal tumour resection that interferes with cancer cell division, slowing tumour recurrence and prolonging life. In GBM patients, this therapeutic modality has demonstrated an increase in median survival from ~16 to ~21 months. This study aimed to assess and decipher the effectiveness of EFT on patient-derived glioblastoma stem cell lines (GSC) as well as on advanced Glioma infiltrating cerebral organoid models (GLICOs). To achieve the project goals, we used a novel In-vitro EFT stimulation apparatus to test stimulation parameters on multiple patient-derived GSC lines as well as GLICO models derived from them. We used advanced microscopy and transcriptomic sequencing to visualize and decipher the causal effect of our stimulation regime on the in-vitro models. We observe a strong effect of our EFT regime on multiple GSC lines as well as on GLICOs when compared to their controls. Transcriptomic analysis showed multitude of differentially expressed mRNAs between the treated and control samples with multiple pathways being dysregulated from nuclear proteins, cell mitosis-related events and activation of non-apoptotic mediated programmed cell death pathways indicating a novel mechanism of action. Advanced causal inference studies using the data also provided novel insights on combinatorial therapies that could enhance the efficacy of the treatment. This study provides novel understanding of the mechanism of EFT with potential for optimised treatment regimens for GBM patients.

CCRG-05. A UNIFYING AND SELF-REINFORCING MECHANISM OF TUMOR IMMORTALITY IN TERT PROMOTER MUTANT CANCERS

Abstract Telomerase Reverse Transcriptase promoter mutations enable tumor cell immortality in millions of cancer patients annually. While prior therapies targeting telomerase lacked tumor selectivity and were poorly tolerated, TERT promoter mutations and their regulation present a unique opportunity for tumor specific reversal of cellular immortality. The two hotspot mutations, G228A and G250A, generate identical de novo E26 transformation specific (ETS) transcription factor binding sites. However, the 28 ETS factors share a similar binding site preference, raising the question of whether all ETS factors can reactivate TERT. In glioblastoma and a few other cancers, we have shown the de novo ETS site along with a nearby native ETS site (ETS-195/200) recruit one ETS factor, the GA-binding protein (GABP) tetramer, to reactivate TERT. This GABP-mediated mechanism may be restricted to specific cancer types and the two hotspot mutations or could be widely relevant to any cancer with a de novo ETS TERT promoter mutation. We have found that the GABP tetramer activates 6 distinct mutant TERT promoters across 16 different cancer types. However, TERT expression is maintained in most tetramer depleted tumor cells as the tetramer is serially replaced by GABP dimers and weakly by a paralogous tetramer complex that increase following disengagement of GABP tetramer-mediated negative feedback loops. Elimination of both the tetramer and dimer reinstates epigenetic repression of TERT, shortens telomeres, and prevents cancer cell division. However, some knockout cancer cells avoid this fate through weak activation of TERT by the paralogous GABP tetramer complex. The compensatory maintenance of TERT expression can be overcome with a dominant negative GABPB1, leading to TERT silencing, evidenced by gene expression and non-invasive metabolic imaging correlates, and telomere shortening in TERT promoter mutant glioblastoma. We are currently investigating the delivery of this dominant negative to tumor cells via clinically approved retroviral replicating vectors.

PATH-38. MOLECULAR CHARACTERIZATION OF GLIOMA TISSUE AFTER TTFIELDS TREATMENT

Abstract INTRODUCTION This study aims to investigate the molecular characterization of glioma tissue after treatment with Tumor Treating Fields (TTFields). TTFields therapy utilizes low-intensity alternating electric fields to disrupt cancer cell division and is approved for treatment of glioblastoma as well as mesothelioma. Understanding the histological, cellular, and epigenetic changes induced by TTFields treatment can help identify biomarkers for TTFields response and elucidate potential resistance mechanisms. MATERIAL AND METHODS Tumor tissue samples (FFPE) were collected from newly diagnosed glioblastoma patients treated with TTFields, including both the primary tumor and matching recurrent tumor samples. Molecular analyses using a glioma-targeted next-generation sequencing (NGS) panel sequencing were performed to assess genetic alterations in both primary and recurrent tumors. RESULTS Genomic analysis of paired samples from 5 patients (4 glioblastoma, one astrocytoma, IDH-mutated WHO grade 4) revealed an identical molecular pattern (TERT mutation) in 2 patients, one patient with a gain of an ATRX mutation in the recurrent tumor, one patient with gain of two EGFR mutations, and the IDH1-mutated astrocytoma showing a loss of a TP53 mutation in the recurrent tumor. CONCLUSION This pilot study provides valuable insights into the molecular characterization of glioma tissue after TTFields treatment. It appears that the spectrum of mutations may vary in a substantial number of TTF-treated tumors. Further validation and larger-scale studies are needed to confirm and expand upon these preliminary results.

QOL-15. THE PIVOTAL TRIDENT STUDY OF TUMOR TREATING FIELDS (TTFIELDS) THERAPY WITH CHEMORADIATION, FOLLOWED BY MAINTENANCE TTFIELDS THERAPY/TEMOZOLOMIDE (TMZ), IN NEWLY DIAGNOSED GLIOBLASTOMA (NDGBM)

Abstract INTRODUCTION: TTFields are electric fields which act to disrupt processes necessary for cancer cell division and tumor progression. TTFields therapy is a noninvasive, locoregional treatment with US FDA approval for ndGBM and recurrent GBM, as well as pleural mesothelioma. TTFields therapy concomitant with maintenance TMZ is standard of care in ndGBM after chemoradiation based on the pivotal EF-14 study (NCT00916409) reporting significant improvements in progression-free survival (PFS) and overall survival (OS) vs TMZ alone. Preclinical and pilot clinical studies subsequently demonstrated that concomitant TTFields therapy also enhanced the therapeutic effect of radiotherapy (RT). TRIAL DESIGN: TRIDENT (EF-32, NCT04471844) is a pivotal multicenter study investigating TTFields therapy (200 kHz) concomitant with RT/TMZ in adult patients with histologically confirmed ndGBM. Eligibility criteria include life expectancy (≥ 3 months) and Karnofsky performance status (≥ 70). Patients (n≈950) stratified by MGMT promoter methylation status and extent of resection are randomly assigned 1:1 to receive TTFields therapy from the first day of chemoradiation (experimental arm), or approximately 4 weeks after completion of chemoradiation (control arm). All patients will receive TTFields therapy uninterrupted for 24 months, or until second disease progression (PFS2). The primary endpoint is median OS and other endpoints include survival rates at 1 and 2 years, PFS (overall, PFS2, and rates at 6 and 12 months), overall radiological response, safety, quality of life and dependence of OS on TTFields dose. The sample size is powered for a hazard ratio of &amp;lt; 0.8 (5% type I error). A stratified log-rank test will be used to test the hypothesis that OS is significantly improved with first line TTFields therapy/RT/TMZ vs RT/TMZ. This global study is open at sites in 11 countries.

Application of Inorganic Nanomaterials for Drug Delivery in Lung Cancer

Lung cancer has been one of the most common cancers in the world for more than 100 years and has taken many people's lives. Many people suffer from lung cancer due to smoking, second-hand smoke, and serious air pollution or pollutant gathering environment. It is necessary to find a good medical solution. Compared with traditional chemotherapy and immunotherapy, targeted therapy has more advantages in the impact on patients and the efficiency of treatment. How to develop targeted therapeutic drugs has become a topic, and nanoscale targeted drugs have become the most appropriate choice. Among them, there are many inorganics suitable for nano-targeted drugs and nano-drug carriers, such as metals, metal oxides, and non-metals. By blocking the genetic material in lung cancer cells, it can avoid the division and differentiation of lung cancer cells, so as to achieve the purpose of treating lung cancer. This review will focus on the above three kinds of nano-targeted drugs, and discuss their therapeutic principles, drug delivery, activation mode, and biotoxicity.

Quantifying assays: inhibition of signalling pathways of cancer.

Inhibiting a signalling pathway concerns controlling the cellular processes of a cancer cell's viability, cell division and death. Assay protocols created to see if the molecular structures of the drugs being tested have the desired inhibition qualities often show great variability across experiments, and it is imperative to diminish the effects of such variability while inferences are drawn. In this paper, we propose the study of experimental data through the lenses of a mathematical model depicting the inhibition mechanism and the activation-inhibition dynamics. The method is exemplified through assay data obtained from an experimental study of the inhibition of the chemokine receptor 4 (CXCR4) and chemokine ligand 12 (CXCL12) signalling pathway of melanoma cells. The quantitative analysis is conducted as a two step process: (i) deriving theoretically from the model the cell viability as a function of time depending on several parameters; (ii) estimating the values of the parameters by using the experimental data. The cell viability is obtained as a function of concentration of the inhibitor and time, thus providing a comprehensive characterization of the potential therapeutic effect of the considered inhibitor, e.g. $IC_{50}$ can be computed for any time point.

An in-silico investigation and network pharmacology based approach to explore the anti-breast-cancer potential of Tecteria coadunata (Wall.) C. Chr.

Uncontrolled cell proliferation is a common definition of cancer. After lung carcinoma, breast neoplasm is the second-most prevalent kind of cancer. The majority of breast cancer cells and healthy breast cells both have receptors for circulating oestrogen and progesterone. In order to promote the development and division of cancer cells, oestrogen and progesterone bind to the receptors and may collaborate with growth factors (such as oncogenes and mutant tumour suppressor genes). As per the literature, Tecteria coadunata (Wall.) C. Chr. has anticancer, antioxidant and anti-inflammatory potential. After the hydroalcoholic extraction of this rhizome, total of 200 phytochemicals were retrieved from HR-LCMS analysis. In this current study, Network pharmacology was carried out to explore the rationale of Tecteria coadunata (Wall.) C. Chr. by using different database using Cytoscape software. The network depicted the interaction of Bioactives with their targets and their association with several disease, especially breast cancer. Tecteria coadunata (Wall.) C. Chr. has offered new relationship with variety of genes and its applications in different types of breast cancers. Further Gene Ontology was carried out and it showed key targets were TP53, BRCA2, PGR and CHEK 2. Further Signalling pathways were also enriched. Flex-X software was used for molecular docking studies, and it verified that Dopaxanthin, Dantrolene and Orotidin shows the highest binding affinities with key targets. Additionally, Pharmacokinetic analysis revealed that all top three lead compounds which follows the Lipinski Rule (Rule of three) without interrupting the conditions of bioavailability with minimal toxicity. Communicated by Ramaswamy H. Sarma

Elucidation of interleukin-19 as a therapeutic target for breast cancer by computational analysis and experimental validation

Interleukin 19 (IL-19) is a cytokine produced by monocytes and belongs to the family of IL-10. The IL-19 protein stimulates fibronectin (FN) expression and assembly, metastasis, and cell division in breast cancer (BC) cells. IL-19, which is connected to breast pathogenesis and has an autocrine action in BC cells, is a key predictor of prognosis for many tumour forms, including breast cancer. Augmented IL-19 expression has been related to poorer clinical outcomes for patients with BC and directly enhances proliferation and migration while also serving as a microenvironment for tumour formation. The main aim of our study was to examine the expression profile, functional role, and prognostic significance of interleukin-19 in BC pathogenesis and also to find out the molecular mechanism of IL-19 in BC. In this work, we used the various computational approach and tools, to evaluate the expression profile and prognostic implication of IL-19 in BC and discover the role of IL-19 in BC pathogenesis. IL-19 was shown to be highly upregulated in BC as compared to other interleukins. Also, its levels were highly overexpressed in liminal BC patients, mostly in 3rd stage groups under the age group of 21–40 years. IL-19 levels were increased in BC and elevated expression of IL-19 was examined to have worse overall survival (OS). The KEGG analysis andgene ontology of IL-19 depict that IL-19 is significantly augmented in cytokine activity and receptor-ligand activity and also in the JAK-STAT signaling pathway. Moreover, IL-19 showed a high correlation with IL20RA, as later is involved with the JAK-STAT signaling pathway. The in-vivo and in-vitro studies have also reflected that upregulation of IL-19 enhances tumor development and affects clinical outcomes in BC patients through several pathways including the JAK TAT signalling pathway. Overall, our study indicates that IL-19 increases tumour growth and that inhibiting it in addition to standard treatments will greatly improve BC patient’s therapeutic responses.

Peptides for Dual Targeting of ErbB1 and ErbB2: Blocking EGFR Cell Signaling Transduction Pathways for Cancer Chemotherapy.

Cancer is one of the most deadly diseases involving dysregulated cell proliferation. Chemotherapeutic drugs have serious drawbacks of nonspecific toxicity and drug resistance. Tyrosine kinases are a significant class of enzymes of protein kinases. The four members of the trans-membrane family of tyrosine kinase receptors known as the human epidermal growth factor receptors (EGFR), ErbB1/HER1, ErbB2/HER2/neu, ErbB3/HER3, and ErbB4/HER4, are overexpressed in many forms of cancer. These receptors are crucial for cell division, invasion, metastasis, angiogenesis, and uncontrolled activation of cancer cells. In this context, an attractive combination of anticancer drug targets is ErbB1 and ErbB2. Numerous cancer types exhibit overexpression of ErbB1 and ErbB2, which is linked to poor prognosis and causes resistance to ErbB1-targeted therapy. Further, it has been reported in recent years that the use of peptides as anticancer agents have the potential to circumvent the drawbacks of the currently used chemotherapeutic drugs. Among them, short peptides have several advantages when compared to small molecules. The present report reviews the importance of tyrosine kinases as targets for cancer, the role of peptides as therapeutic agents, and the investigations that have been carried out by earlier workers for targeting both ErbB1 and ErbB2 using therapeutic peptides.

Cellular, Biophysical and in Silico Binding Study of β-Estradiol-6-one 6- (O-carboxy methyl Oxime) with Tubulin in Search of Antimitotic Derivative of 2-Methoxy Estradiol.

The tubulin-microtubule system is a major target for a variety of small molecules which can interfere in cell cycle progression. Therefore, it serves as a prospective to control the incessant division of cancer cells. To identify novel inhibitors of the tubulin-microtubule system, a group of estrogen derivatives has been tested with tubulin as a target since literature surveys portray coveted behaviour from the same. Out of them, β-Estradiol-6-one 6- (O-carboxy methyl Oxime) abbreviated as Oxime, disrupts the cytoskeleton network and induces apoptosis with nuclei fragmentation. It has been revealed from the work that Oxime targets the colchicine binding site and binds tubulin in an entropy-driven manner. This suggests that structural variation might play a key role in modulating the anti-mitotic role of estrogen derivatives. Our work reveals that Oxime might serve as a lead molecule to nurture anti-cancer research, having the potential for recovery of the vast cancer population.

PENAMBATAN MOLEKUL SIANIDIN Ipomoea batatas L. SEBAGAI INHIBITOR HUMAN EPIDERMAL RECEPTOR 2 (HER-2) PADA KANKER PAYUDARA

HER-2 (Human Epidermal Receptor 2) is a receptor on the cell surface that affects cell proliferation, this receptor support controls healthy cell growth,divide and repair cells when they are damaged. When HER-2 had overexpressed will not controlled cancer cell division. Overexpressed HER-2 could inhibited with cyanidine compound flavonoid derivatives from Ipomoea batatas L. (purple sweet potato). This study aim to measured potential of Ipomoea batatas L. cyanidin compounds in inhibited overexpressed HER-2 using computational chemistry methods specifically is molecular docking. Stages of molecular docking receptor HER-2 is ligand set up with drew structure cyanidine with Marvin Sketch then optimized by Autodock Tools and Discovery Studio, lapatinib as a comparison ligand downloaded on PubChem website. HER-2 (3PP0) prepared with downloaded on Protein Data Bank website then optimized by Autodock Tools. Molecular Docking validated with docked protein target with native ligand by PyMOL to secured RMSD (root mean square deviation). Protein Targeted HER-2 docked with test ligand cyanidine, lapatinib, and native ligand by Autodock VINA, analysed energy bond and amino acid residue interactions by LigPlot+. Cyanidine had potency inhibitor of receptor HER-2 overexpressed from docked is had bound energy -7.1 ccal/mol up to -8.6 ccal/mol wich is also had amino acid residue interactions Met 801 equal to native ligand

Abstract 5989: Targeting RRM2 &amp; cell cycle for breast cancer treatment

Abstract Endocrine treatment is an effective first-line therapy for targeting ER positive, HER2 negative breast cancer. But success is limited by development of acquired resistance due to long-term therapy. Cyclin D1 and cyclin dependent kinase 4/6 (CDK4/6) complex causes phosphorylation and subsequent inactivation of retinoblastoma (Rb) tumor suppressor protein which promotes progression of the cell cycle from G1 to S phase. This observation led the development of the first CDK4/6 inhibitor palbociclib (Ibrance; Pfizer) which induces cell cycle arrest at G1 phase in cancer cells. Owing to intrinsic and acquired drug resistance development, success is limited despite promising clinical outcomes. This situation necessitates the development of potential combination strategies to overcome drug resistance. Ribonucleotide reductase (RR) is a rate limiting enzyme in DNA synthesis consisting of two subunits RRM1 and RRM2. Didox inhibits ribonucleotide reductase subunit 2 (RRM2) which ultimately blocks DNA synthesis. Combination of didox with palbociclib is a potential strategy to target ER positive and ER negative/triple negative breast cancer. Previously, we have shown that didox (DDX) can significantly halt malignant breast cancer cell division in combination with the anthracycline drug doxorubicin by targeting RRM2, mutant p53 and NFkB regulatory proteins. Recently, we confirmed that didox in combination with palbociclib significantly lowers ER positive and ER negative breast cancer and their palbociclib resistant counterparts’ growth compared to no treatment or palbociclib treatment. We also confirmed that ER positive MCF7 and ER negative MDA-MB-468 parental breast cancer cells IC50 of palbociclib drug is lower than their palbociclib resistant counterparts. Here, we are reporting that didox decreases cell cycle proteins alone or in combination with palbociclib in ER positive MCF7 and ER negative MDA-MB-468 parental and palbociclib resistant breast cancer cells. This finding opens a novel approach for targeting both ER positive as well as ER negative breast cancer treatment. We are also reporting that didox treatment modulates cyclin D1 (CCND1) and RRM2 expression in MCF7 and MDA-MB-468 breast cancer along with their palbociclib resistant counterparts. Additionally, we observed that didox alone or in combination with palbociclib alters the cell cycle of MCF7 and MDA-MB-468 parental and palbociclib resistant breast cancer cells. Our data presents a novel and promising approach for the treatment of ER positive and ER negative breast cancer that involves inhibition of RRM2 and cell cycle that merits further clinical investigation in animal and human models. Citation Format: Nahid Sultana, Howard L. Elford, Jesika S. Faridi. Targeting RRM2 &amp; cell cycle for breast cancer treatment. [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 5989.

Abstract 6715: CLIC1 and CLIC4 ion channels as bioelectric targets for tumor treating fields in pediatric high-grade glioma

Abstract Background: High Grade Glioma is a lethal brain cancer with an average survival of 14 months. Due to the scarcity of effective treatment, and aggressive nature of the tumor, pediatric High Grade Glioma (pHGG) forms a leading cause of CNS cancer-related death in children. Optune™ is a non-invasive therapy that uses alternating electric fields - coined Tumor Treating Fields (TTFields) - to disrupt cancer cell division. Increasing evidence shows that ion channels not only regulate electrical signaling of excitable cells, but also play a crucial role in the development and progression of brain tumors. Ion channels are essential in cell cycle control, invasion and migration of cancer cells and therefore present as valuable therapeutic targets. Methods: CLIC channel expression patterns were identified via multivariate analysis of in-house and publicly available data sets and RNA sequencing of in-house patient tissues. siRNA depletion of CLIC1 and CLIC4 was assessed using cell cycle, clonogenic, migration and proliferation assays, alone and concomitant with TTFields and or CLIC channel inhibitors, IAA94 or metformin. Whole transcriptome gene expression analysis (Human Clariom™ Array) of pHGG cells treated with TTFields was carried out. Whole cell and cell attached patch clamp protocols were used to assess ion channel activity in pHGG and normal astrocytes. Results: Clinical correlation determined that CLIC4 and CLIC1 deficiency was associated with increased overall survival (p=&amp;lt;0.03), with siRNA depletion propagating a reduction in the proliferation, migration and invasion of pHGG cell lines associated with cell cycle arrest. Furthermore, CLIC1 and CLIC4 deficiency exacerbated the killing capacity of TTFields, reducing clonogenic and proliferative capabilities. Concomitant application of either IAA94 or metformin, and TTFields increased the sensitivity of cells to TTFields. Whole transcriptome gene expression analysis of pHGG cell lines treated with TTFields and found that cells treated with TTFields exhibited a down-regulation in CLIC1 and CLIC4 compared to untreated cells. Furthermore, neurophysiological patch clamp experiments revealed increasingly depolarized membrane potential readings in pHGG cell lines compared to normal astrocytes, with unique ion channel activity (including chloride channel function) observed and the potential for further pharmacological exploitation. Conclusions: These data provide rationale that genetic, electrical, and pharmacological manipulation of ion channels can reduce the capacity of childhood brain tumors to proliferate and invade. CLIC channels may be a suitable target to enhance the treatment efficacy of TTFields and help optimize this non-invasive therapy in pediatric HGG patients. Citation Format: Michaela Griffin, Paul Smith, Raheela Khan, Surajit Basu, Stuart James Smith. CLIC1 and CLIC4 ion channels as bioelectric targets for tumor treating fields in pediatric high-grade glioma [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 6715.

СРАВНИТЕЛЬНЫЙ АНАЛИЗ СОСТАВА И КОЛИЧЕСТВЕННОГО СОДЕРЖАНИЯ АНТОЦИАНОВ В ПЛОДАХ КЛЮКВЫ ОБЫКНОВЕННОЙ (VACCINIUM OXYCOCCUS L.) И КРУПНОПЛОДНОЙ (VACCINIUM MACROCARPON AIT.)

The authors carry out a comparative analysis of the fruits of the common cranberry, used in the Russian Federation as food and medicinal raw materials, and the fruits of the large-fruited cranberry, which is the most cultivated species, the cultivation of which began in the 19th century in Canada and the United States, and in Russia has been widely spread since 1980. An important group of biologically active substances in cranberry fruits is undoubtedly anthocyanins [5], which have a pronounced antioxidant activity, help strengthen the walls of blood vessels, stimulate the processes of protein biosynthesis, and have a stimulating effect on reparative processes. In vitro and in vivo experiments revealed that anthocyanin pigments can significantly reduce the rate of cancer cell division. HPLC method established the presence of anthocyanins in the fruits of common and large-fruited cranberries. In the composition of the anthocyanin fraction of all the studied samples, peonidin-3-arabinoside was identified, the presence of which makes it possible to reliably identify cranberry fruits and their processed products. The total amount of detected anthocyanin pigments ranges from 0.17% (1.31% by weight of absolutely dry raw materials) in fruits of fresh common cranberries to 0.27% (1.64% by weight of absolutely dry raw materials) in fruits of large-fruited cranberries of the Mack variety -Farlin The observed differences can be associated with both ecological and geographical factors, i.e. the difference caused by the harvesting of raw materials in different places where cranberries grow, and the season of harvesting fruits for analysis, which undoubtedly requires additional study of cranberry fruits from different regions of harvesting and harvesting time and will be continued in subsequent studies. The authors also found that during the preservation of cranberries by freezing, the content of anthocyanins decreases, which makes it possible to recommend the maximum processing of fresh raw materials.

Abstract P1-13-01: Targeting RRM2 for the treatment of palbociclib resistant breast cancer

Abstract Hormone receptor (HR) positive breast cancer (BC) is a prevalent disease accounting for almost 2 million new cases globally. Almost 70-80% of breast cancer patients are women with a positive score for the estrogen receptor (ER). HR positive BCs of all stages are selectively treated with endocrine therapy targeting ER activity. Intrinsic and acquired resistance are common phenomena, impacting patient outcomes negatively. Palbociclib is an FDA approved checkpoint inhibitor for the treatment of HR+/HER2- breast cancer in combination with aromatase inhibitors or selective estrogen receptor degraders. However, patients responding well to the therapy in the beginning, lose sensitivity to palbociclib with time. Ribonucleotide reductase (RR) is a rate limiting enzyme in DNA synthesis consisting of two subunits RRM1 and RRM2. In our previous study, we have shown that RRM2 is upregulated in ER positive tamoxifen resistant BC. Previously, we have also reported that didox (DDX) which is a unique RRM2 enzyme inhibitor can significantly halt malignant breast cancer cell division in combination with an anthracycline drug doxorubicin by targeting RRM2, mutant p53 and NFkB regulatory proteins. In this study, we target palbociclib resistant BC with DDX to circumvent palbociclib resistance. For this purpose, we have developed palbociclib resistant ER positive MCF7 and ER negative MDA-MB-468 breast cancer cell lines. Here, we report morphological changes in parental as compared to palbociclib resistant cells after treatment with palbociclib alone, DDX alone or with palbociclib as compared to non-treated cells. Cell proliferation studies confirm the synergistic combinatorial effect of palbociclib and DDX compared to palbociclib or DDX alone. We also report that DDX alone or in combination with palbociclib downregulates cell cycle and NFkB proteins in palbociclib resistant ER positive MCF7 and ER negative MBA-MB-468 breast cancer cell lines. Citation Format: Nahid Sultana, Howard L. Elford, Jesika S. Faridi. Targeting RRM2 for the treatment of palbociclib resistant breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P1-13-01.