Affect Cancer Cell Research Articles

Flavokawain A is a natural inhibitor of PRMT5 in bladder cancer

BackgroundProtein arginine methyltransferases (PRMTs) regulate protein biological activity by modulating arginine methylation in cancer and are increasingly recognized as potential drug targets. Inhibitors targeting PRMTs are currently in the early phases of clinical trials and more candidate drugs are needed. Flavokawain A (FKA), extracted from kava plant, has been recognized as a potential chemotherapy drug in bladder cancer (BC), but its action mechanism remains unclear.MethodsWe first determined the role of a type II PRMT, PRMT5, in BC tissue samples and performed cytological experiments. We then utilized bioinformatics tools, including computational simulation, virtual screening, molecular docking, and energy analysis, to identify the potential use of PRMT5 inhibitors for BC treatment. In vitro and in vivo co-IP and mutation assays were performed to elucidate the molecular mechanism of PRMT5 inhibitor. Pharmacology experiments like bio-layer interferometry, CETSA, and pull-down assays were further used to provide direct evidence of the complex binding process.ResultsAmong PRMTs, PRMT5 was identified as a therapeutic target for BC. PRMT5 expression in BC was correlated with poor prognosis and manipulating its expression could affect cancer cell growth. Through screening and extensive experimental validation, we recognized that a natural product, FKA, was a small new inhibitor molecule for PRMT5. We noticed that the product could inhibit the action of BC, in vitro and in vivo, by inhibiting PRMT5. We further demonstrated that FKA blocks the symmetric arginine dimethylation of histone H2A and H4 by binding to Y304 and F580 of PRMT5.ConclusionsIn summary, our research strongly suggests that PRMT5 is a potential epigenetic therapeutic target in bladder cancer, and that FKA can be used as a targeted inhibitor of PRMT5 for the treatment of bladder cancer.

Phytochemicals in Inhibition of Prostate Cancer: Evidence from Molecular Mechanisms Studies.

Prostate cancer is one of the leading causes of death for men worldwide. The development of resistance, toxicity, and side effects of conventional therapies have made prostate cancer treatment become more intensive and aggressive. Many phytochemicals isolated from plants have shown to be tumor cytotoxic. In vitro laboratory studies have revealed that natural compounds can affect cancer cell proliferation by modulating many crucial cellular signaling pathways frequently dysregulated in prostate cancer. A multitude of natural compounds have been found to induce cell cycle arrest, promote apoptosis, inhibit cancer cell growth, and suppress angiogenesis. In addition, combinatorial use of natural compounds with hormone and/or chemotherapeutic drugs seems to be a promising strategy to enhance the therapeutic effect in a less toxic manner, as suggested by pre-clinical studies. In this context, we systematically reviewed the currently available literature of naturally occurring compounds isolated from vegetables, fruits, teas, and herbs, with their relevant mechanisms of action in prostate cancer. As there is increasing data on how phytochemicals interfere with diverse molecular pathways in prostate cancer, this review discusses and emphasizes the implicated molecular pathways of cell proliferation, cell cycle control, apoptosis, and autophagy as important processes that control tumor angiogenesis, invasion, and metastasis. In conclusion, the elucidation of the natural compounds’ chemical structure-based anti-cancer mechanisms will facilitate drug development and the optimization of drug combinations. Phytochemicals, as anti-cancer agents in the treatment of prostate cancer, can have significant health benefits for humans.

Hsp70-Bag3 Module Regulates Macrophage Motility and Tumor Infiltration via Transcription Factor LITAF and CSF1.

Simple SummaryPatients’ normal cells, such as lymphocytes, fibroblasts, or macrophages, can either suppress or facilitate tumor growth. Macrophages can infiltrate tumors and secrete molecules that enhance the proliferation of cancer cells and their invasion into neighboring tissues and blood. Here, we investigated the mechanism of action of a novel small molecule that suppresses the infiltration of macrophages into tumors and demonstrates potent anticancer activity. We identified the entire pathway that links the intracellular protein Hsp70, which is inhibited by this small molecule, with the macrophage motility system. This study will lay the basis for a novel approach to cancer treatment via targeting tumor-associated macrophages.The molecular chaperone Hsp70 has been implicated in multiple stages of cancer development. In these processes, a co-chaperone Bag3 links Hsp70 with signaling pathways that control cancer development. Recently, we showed that besides affecting cancer cells, Hsp70 can also regulate the motility of macrophages and their tumor infiltration. However, the mechanisms of these effects have not been explored. Here, we demonstrated that the Hsp70-bound co-chaperone Bag3 associates with a transcription factor LITAF that can regulate the expression of inflammatory cytokines and chemokines in macrophages. Via this interaction, the Hsp70–Bag3 complex regulates expression levels of LITAF by controlling its proteasome-dependent and chaperone-mediated autophagy-dependent degradation. In turn, LITAF regulates the expression of the major chemokine CSF1, and adding this chemokine to the culture medium reversed the effects of Bag3 or LITAF silencing on the macrophage motility. Together, these findings uncover the Hsp70–Bag3–LITAF–CSF1 pathway that controls macrophage motility and tumor infiltration.

IRE1α overexpression in malignant cells limits tumor progression by inducing an anti-cancer immune response

ABSTRACT IRE1α is one of the three ER transmembrane transducers of the Unfolded Protein Response (UPR) activated under endoplasmic reticulum (ER) stress. IRE1α activation has a dual role in cancer as it may be either pro- or anti-tumoral depending on the studied models. Here, we describe the discovery that exogenous expression of IRE1α, resulting in IRE1α auto-activation, did not affect cancer cell proliferation in vitro but resulted in a tumor-suppressive phenotype in syngeneic immunocompetent mice. We found that exogenous expression of IRE1α in murine colorectal and Lewis lung carcinoma cells impaired tumor growth when syngeneic tumor cells were subcutaneously implanted in immunocompetent mice but not in immunodeficient mice. Mechanistically, the in vivo tumor-suppressive effect of overexpressing IRE1α in tumor cells was associated with IRE1α RNAse activity driving both XBP1 mRNA splicing and regulated IRE1-dependent decay of RNA (RIDD). We showed that the tumor-suppressive phenotype upon IRE1α overexpression was characterized by the induction of apoptosis in tumor cells along with an enhanced adaptive anti-cancer immunosurveillance. Hence, our work indicates that IRE1α overexpression and/or activation in tumor cells can limit tumor growth in immunocompetent mice. This finding might point toward the need of adjusting the use of IRE1α inhibitors in cancer treatments based on the predominant outcome of the RNAse activity of IRE1α.

ADAR2 Protein Is Associated with Overall Survival in GBM Patients and Its Decrease Triggers the Anchorage-Independent Cell Growth Signature.

Background: Epitranscriptomic mechanisms, such as A-to-I RNA editing mediated by ADAR deaminases, contribute to cancer heterogeneity and patients’ stratification. ADAR enzymes can change the sequence, structure, and expression of several RNAs, affecting cancer cell behavior. In glioblastoma, an overall decrease in ADAR2 RNA level/activity has been reported. However, no data on ADAR2 protein levels in GBM patient tissues are available; and most data are based on ADARs overexpression experiments. Methods: We performed IHC analysis on GBM tissues and correlated ADAR2 levels and patients’ overall survival. We silenced ADAR2 in GBM cells, studied cell behavior, and performed a gene expression/editing analysis. Results: GBM tissues do not all show a low/no ADAR2 level, as expected by previous studies. Although, different amounts of ADAR2 protein were observed in different patients, with a low level correlating with a poor patient outcome. Indeed, reducing the endogenous ADAR2 protein in GBM cells promotes cell proliferation and migration and changes the cell’s program to an anchorage-independent growth mode. In addition, deep-seq data and bioinformatics analysis indicated multiple RNAs are differently expressed/edited upon siADAR2. Conclusion: ADAR2 protein is an important deaminase in GBM and its amount correlates with patient prognosis.

α/β-Hydrolase Domain-Containing 6 (ABHD6)- A Multifunctional Lipid Hydrolase.

α/β-hydrolase domain-containing 6 (ABHD6) belongs to the α/β-hydrolase fold superfamily and was originally discovered in a functional proteomic approach designed to discover monoacylglycerol (MAG) hydrolases in the mouse brain degrading the endocannabinoid 2-arachidonoylglycerol. Subsequent studies confirmed that ABHD6 acts as an MAG hydrolase regulating cannabinoid receptor-dependent and -independent signaling processes. The enzyme was identified as a negative modulator of insulin secretion and regulator of energy metabolism affecting the pathogenesis of obesity and metabolic syndrome. It has been implicated in the metabolism of the lysosomal co-factor bis(monoacylglycerol)phosphate and in the surface delivery of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptors. Finally, ABHD6 was shown to affect cancer cell lipid metabolism and tumor malignancy. Here, we provide new insights into the experimentally derived crystal structure of ABHD6 and its possible orientation in biological membranes, and discuss ABHD6′s functions in health and disease.

A Tumor Accelerator Based on Multicomponent Bone Scaffolds and Cancer Cell Homing.

Bone tissue attracts cancer cell homing biologically, mechanically, or chemically. It is difficult and time consuming to identify their complex cross-talk using existed methods. In this study, a multi-component bone matrix was fabricated using gelatin, hydroxyapatite (HAp), and epidermal growth factor (EGF) as raw materials to investigate how “acellular” bone matrix affects cancer cell homing in bone. Then, EGF-responsive cancer cells were cultured with the scaffold in a dynamical bioreactor. For different culture periods, the effects of HAp, gelatin, and EGF on the cell adhesion, proliferation, 3D growth, and migration of cancer were evaluated. The results indicated that a small amount of calcium ion released from the scaffolds accelerated cancer MDA-MB-231 adhesion on the surface of inner pores. Moreover, degradable gelatin key caused cancer cell growth on the scaffold surface to turn into a 3D aggregation. Despite this, the formation of cancer spheroids was slow, and required 14 days of dynamic culture. Thankfully, EGF promoted cancer cell adhesion, proliferation, and migration, and cancer spheroids were observed only after 3-day culture. We concluded that the combination of the multiple components in this scaffold allows cancer cells to meet multiple requirements of cancer dynamic progression.

Microbiota-driven mechanisms at different stages of cancer development

SummaryA myriad of microbes living together with the host constitutes the microbiota, and the microbiota exerts very diverse functions in the regulation of host physiology. Microbiota regulates cancer initiation, progression, metastasis, and responses to therapy. Here we review known pro-tumorigenic and anti-tumorigenic functions of microbiota, and mechanisms of how microbes can shape tumor microenvironment and affect cancer cells as well as activation and functionality of immune and stromal cells within the tumor. While some of these mechanisms are distal, often distinct members of microbiota travel with and establish colonization with the tumors in the distant organs. We further briefly describe recent findings regarding microbiota composition in metastasis and highlight important future directions and considerations for the manipulation of microbiota for cancer treatment.

Hybrid Molecules Containing Naphthoquinone and Quinolinedione Scaffolds as Antineoplastic Agents.

In recent decades, molecular hybridization has proven to be an efficient tool for obtaining new synthetic molecules to treat different diseases. Based on the core idea of covalently combining at least two pharmacophore fragments present in different drugs and/or bioactive molecules, the new hybrids have shown advantages when compared with the compounds of origin. Hybridization could be successfully applied to anticancer drug discovery, where efforts are underway to develop novel therapeutics which are safer and more effective than those currently in use. Molecules presenting naphthoquinone moieties are involved in redox processes and in other molecular mechanisms affecting cancer cells. Naphthoquinones have been shown to inhibit cancer cell growth and are considered privileged structures and useful templates in the design of hybrids. The present work aims at summarizing the current knowledge on antitumor hybrids built using 1,4- and 1,2-naphthoquinone (present in natural compounds as lawsone, napabucasin, plumbagin, lapachol, α-lapachone, and β -lapachone), and the related quinolone- and isoquinolinedione scaffolds reported in the literature up to 2021. In detail, the design and synthetic approaches adopted to produce the reported compounds are highlighted, the structural fragments considered in hybridization and their biological activities are described, and the structure–activity relationships and the computational analyses applied are underlined.

СРАВНИТЕЛЬНАЯ ХАРАКТЕРИСТИКА ЛЕКАРСТВЕННЫХ ПРЕПАРАТОВ РОССИЙСКОГО ПРОИЗВОДСТВА ДЛЯ ФОТОДИНАМИЧЕСКОЙ ТЕРАПИИ

Abstract. Currently the using of photodynamic therapy (PDT) is expanding in clinical practice. This type of treatment is used not only in oncology, but also in such areas as dentistry, cardiology, ophthalmology, surgery, dermatology, otolaryngology and cosmetology. An urgent task facing clinicians using photodynamic methods is to choose the optimal photosensitizer (PS) and modes of its photoactivation, the result of which will lead to the necessary therapeutic effect. Conditionally, the PS is divided into three generations. Hematoporphyrin derivatives are first-generation PS. The most well-known and widely used is Photofrin. The second generation of PS is represented by chlorines, phthalocyanines, benzoporphyrins, 5-aminolevulinic acid. Foscan is the strongest PS of the second generation. A serious drawback in the first and second generation of PS: their selective accumulation in the tumor cell is impossible. The third generation of PS is the means of the second generation associated with a carrier that provides minimal accumulation in healthy tissues with high specificity to the affected cancer cell. To create a complete picture of the main aspects of the application, it is necessary to know the physicochemical and commodity characteristics of such drugs. The authors of the article reviewed the range of PS registered in the Russian Federation and used in clinical practice. Their attitude to the pharmacotherapeutic group, distribution according to the anatomical and therapeutic classification are shown, physicochemical properties, indications, side effects are characterized. The classifications are compared, the advantages and disadvantages of photosensitizers used for the treatment of malignant tumors and other diseases by PDT are considered. A classification of PS is proposed based on their application in combination with physicochemical properties. It is indicated that the main active substances of Russian-made medicines are derivatives of porphyrins and aminolevulinic acid. Thus, at present, the range of PS allows to significantly expand the possible areas of application and increase the effectiveness of PDT in various diseases, but also to continue the search for the ideal drug for this type of therapy.

Cholesterol homeostasis and cancer: a new perspective on the low-density lipoprotein receptor.

Disturbance of cholesterol homeostasis is considered as one of the manifestations of cancer. Cholesterol plays an essential role in the pleiotropic functions of cancer cells, including mediating membrane trafficking, intracellular signal transduction, and production of hormones and steroids. As a single transmembrane receptor, the low-density lipoprotein receptor (LDLR) can participate in intracellular cholesterol uptake and regulate cholesterol homeostasis. It has recently been found that LDLR is aberrantly expressed in a broad range of cancers, including colon cancer, prostate cancer, lung cancer, breast cancer and liver cancer. LDLR has also been found to be involved in various signaling pathways, such as the MAPK, NF-κB and PI3K/Akt signaling pathways, which affect cancer cells and their surrounding microenvironment. Moreover, LDLR may serve as an independent prognostic factor for lung cancer, breast cancer and pancreatic cancer, and is closely related to the survival of cancer patients. However, the role of LDLR in some cancers, such as prostate cancer, remains controversial. This may be due to the lack of normal feedback regulation of LDLR expression in cancer cells and the severe imbalance between LDLR-mediated cholesterol uptake and de novo biosynthesis of cholesterol. The imbalance of cholesterol homeostasis caused by abnormal LDLR expression provides new therapeutic opportunities for cancer. LDLR interferes with the occurrence and development of cancer by modulating cholesterol homeostasis and may become a novel target for the development of anti-cancer drugs. Herein, we systematically review the contribution of LDLR to cancer progression, especially its dysregulation and underlying mechanism in various malignancies. Besides, potential targeting and immunotherapeutic options are proposed.

The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition.

Serum-containing medium is widely used to support cell attachment, stable growth and serial passaging of various cancer cell lines. However, the presence of cholesterols and lipids in serum greatly hinders the analysis of the effects of cholesterol depletion on cells in culture. In this study, we developed a defined serum-free culture condition accessible to a variety of different types of adherent cancer cells. We tested different factors that are considered essential for cell culture and various extracellular matrix for plate coating, and found cells cultured in Dulbecco's Modified Eagle's Medium (DMEM) basal media supplemented with Albumin (BSA) and insulin-transferrin-selenium-ethanolamine (ITS-X) on fibronectin-precoated plate (called as "DA-X condition") showed comparable proliferation and survival to those in a serum-containing medium. Interestingly, we observed that DA-X condition could be adapted to a wide variety of adherent cancer cell lines, which enabled the analysis of how cholesterol depletion affected cancer cells in culture. Mechanistically, we found the beneficial effects of the DA-X condition in part can be attributed to the appropriate level of membrane cholesterol, and fibronectin-mediated signaling plays an important role in the suppression of cholesterol production.

A Hybrid Model Particle Swarm Optimization Based Mammogram Classification Using Kernel Support Vector Machine

Identifying affected cancer cells in women’s breasts is mammogram, which is the major issue in the field of medicine all over the world. In order to raise the endurance of patients, it is most essential to identify the issue as early as possible. It also helps them to inflate the different options for treatment. With the new dramatic development in computation, machine learning made a revolution with dataset includes huge volume of breast images which could assist in recognizing malignant tumor with better diagnostics. Digital mammography images are taken, in that the x-ray images are read and stored in computer such that data can be easily enhanced and classified for further action. A novel approach is proposed in this paper to diagnose cancer affected cells with a good accuracy rate. Classification of mammogram with hybrid model includes feature extraction, various kinds of features are extorted from the intensity mammogram. A Particle Swarm Optimization optimizer is used in this paper which selects the features, and kernel-based Support Vector Machine classifier classifies the cancer lump from the taken mammogram metaphors. The exactness of a specific model can be assessed by the level of right forecasts made by the model.

Role of metformin in prevention and prognosis of breast cancer

Dear Editor, Metformin is a hypoglycemic drug of the class biguanides used as the first-line treatment for type 2 diabetes mellitus (T2DM). The drug acts by inhibiting hepatic gluconeogenesis, decreasing intestinal absorption of glucose and circulating insulin. It has also been suggested that the drug indirectly increases insulin sensitivity by increasing peripheral glucose utilization. Data from the World Health Organization (WHO) shows that breast cancer (BC) is the most prevalent cancer in women, with an incidence of 1.7 million cases in 2012 (25 % of all female cancers). It is the fifth leading cause of death from cancer overall.1 A clinical trial conducted in 2019 elaborated that metformin altered host metabolism proving to be a substitute for a fasting-like mechanism that affects cancer cells' metabolism and growth environment of cancer cells including an increase in beta-OH-butyric acid (BHBA).2 It was further corroborated by a research that found an inverse relationship between metformin and BC. This is accomplished by boosting the activity of AMP-dependent protein kinase, which suppresses the mTOR-associated oncogenic signalling pathway. This induces S phase arrest and apoptosis in triple-negative breast cancer cells. Data from 18,527 participants brought about 2,680 participants who used metformin at least three months before the diagnosis of BC. A multivariate study showed that this was negatively associated with BC risk (0.821, 0.726-0.912, p=0.002).3 A favourable response on disease-free survival and overall survival in BC patients with T2DM. The number of CD8+ tumour-infiltrating lymphocytes (TILS) has been increased with the use of metformin engendering anti-carcinogenic effects.4 In the last 15 years, we have seen a drastic change in the demographics of BC, with an estimated increase of 70.7 to 130.6% in 2020 and 2025 relative to 2015 in the 30-34 age group in Pakistan.5 There has been a significant increase in T2DM along with increased use of metformin for its treatment which thus raises a question about the efficacy of the drug in a neoadjuvant setting for BC on Pakistani society. Further research, especially clinical trials, needs to be conducted in Pakistan including but not limited to a correlation of metformin use and incidence of BC.

High-Intensity Aerobic Exercise Suppresses Cancer Growth by Regulating Skeletal Muscle-Derived Oncogenes and Tumor Suppressors.

High-intensity aerobic exercise (90% of the maximal heart rate) can effectively suppress cancer cell proliferation in vivo. However, the molecular effects of exercise and its relevance to cancer prevention remain uninvestigated. In this study, mice with colorectal cancer were subjected to high-intensity aerobic exercise, and mRNA-seq analysis was performed on the heart, lungs, and skeletal muscle tissues to analyze the genome-wide molecular effects of exercise. The skeletal muscle-derived genes with exercise-dependent differential expression were further evaluated for their effects on colorectal cancer cell viability. Compared to the results obtained for the control groups (healthy and cancer with no exercise), the regular and high-intensity aerobic physical activity in the mice produced positive results in comprehensive parameters (i.e., food intake, weight gain, and survival rate). A heatmap of differentially expressed genes revealed markedly different gene expression patterns among the groups. RNA-seq analysis of 23,282 genes expressed in the skeletal muscle yielded several anticancer effector genes (e.g., Trim63, Fos, Col1a1, and Six2). Knockdown and overexpression of selected anticancer genes repressed CT26 murine colorectal carcinoma cell proliferation by 20% (p < 0.05). Our findings, based on the aerobic exercise cancer mouse model, suggest that high-intensity aerobic exercise results in a comprehensive change in the expression patterns of genes, particularly those that can affect cancer cell viability. Such an approach may identify key exercise-regulated genes that can help the body combat cancer.

Cytoskeleton disruption by the metabolic inhibitor 3-bromopyruvate: implications in cancer therapy.

The small molecule 3-bromopyruvate (3BP), is an anticancer molecule that acts by hindering glycolysis and mitochondrial function leading to energy depletion and consequently, to cell death. In this work we have focused on understanding how the glycolytic inhibition affects cancer cell structural features. We showed that 3BP leads to a drastic decrease in the levels of β-actin and α-tubulin followed by disorganization and shrinkage of the cytoskeleton in breast cancer cells. 3BP inhibits cell migration and colony formation independently of the activity of metalloproteinases. To disclose if these structural alterations occurred prior to 3BP toxic effect, non-toxic concentrations of 3BP were used and we could observe that 3BP was able to inhibit energy production and induce loss of β-actin and α-tubulin proteins. This was accompanied with alterations in cytoskeleton organization and an increase in E-cadherin levels which may indicate a decrease in cancer cells aggressiveness. In this study we demonstrate that 3BP glycolytic inhibition of breast cancer cells is accompanied by cytoskeleton disruption and consequently loss of migration ability, suggesting that 3BP can potentially be explored for metastatic breast cancer therapy.

Selective cell cycle arrest in glioblastoma cell lines by quantum molecular resonance alone or in combination with temozolomide

BackgroundGlioblastoma is the most aggressive form of brain cancer, characterised by high proliferation rates and cell invasiveness. Despite advances in surgery and radio-chemotherapy, patients continue to have poor prognoses, with a survival rate of 14–15 months. Thus, new therapeutic strategies are needed. Non-ionising electromagnetic fields represent an emerging option given the potential advantages of safety, low toxicity and the possibility to be combined with other therapies.MethodsHere, the anticancer activity of quantum molecular resonance (QMR) was investigated. For this purpose, three glioblastoma cell lines were tested, and the QMR effect was evaluated on cancer cell proliferation rate and aggressiveness. To clarify the QMR mechanism of action, the proteomic asset after stimulation was delineated. Mesenchymal stromal cells and astrocytes were used as healthy controls.ResultsQMR affected cancer cell proliferation, inducing a significant arrest of cell cycle progression and reducing cancer tumorigenicity. These parameters were not altered in healthy control cells. Proteomic analysis suggested that QMR acts not only on DNA replication but also on the machinery involved in the mitotic spindle assembly and chromosome segregation. Moreover, in a combined therapy assessment, QMR significantly enhanced temozolomide efficacy.ConclusionsQMR technology appears to be a promising tool for glioblastoma treatment.

Abstract 2693: The vitamin D3 metabolite, 24R,25(OH)3D3 is differentially tumorigenic in ER+ and ER- laryngeal cancer cells in vitro

Abstract Due to the high vitamin D3 levels associated with improved prognosis (but not incidence) in post-menopausal breast cancer patients with ER+ tumors, vitamin D3 supplementation has been used as potential therapeutics. 24R,25(OH)2D3, an active metabolite of vitamin D3, is known to protect cartilage cells through a membrane-initiated, p53-mediated pathway, suggesting that 24R,25(OH)2D3 may affect cancer cells through a similar mechanism. Our lab has shown that the effect of 24R,25(OH)2D3 on breast cancer cell growth is highly dependent on the expression levels of ERα isoforms, specifically ERα66. Understanding the mechanisms by which 24R,25(OH)2D3 affects tumor growth in breast cancer will aid in determining its therapeutic potential in another estrogen responsive cancer, laryngeal cancer. The goal of this study was to examine the effects of 24R,25(OH)2D3 on proliferation, apoptosis, and epithelial to mesenchymal transition (EMT) in laryngeal cancer cell lines. To assess the effect of 24R,25(OH)2D3 on cell growth, UM-SCC-12 (ERα66+) and UM-SCC-11A (ERα66-) cell lines were cultured on 96 well plates until 80% confluence. Cells were serum starved for 24 hours pretreatment and then treated with full media containing either vehicle, or 24R,25(OH)2D3 (10-9M, 10-8M, or 10-7M) for 15 minutes, at which time the media were replaced with fresh media. After 20 hours, cells were harvested for proliferation by EDU assay. For other experiments cells were treated with either vehicle, or 24R,25(OH)2D3 (10-9M, 10-8M, or 10-7M) for 15 minutes and after 24 hours, cells were assessed for apoptosis (TUNEL, P53) and metastasis (OPG/RANKL). 12 hours after treatment, cells were harvested for PCR and examined for gene expression of apoptotic markers (BAX, BCL2) and various metastatic markers (SNAI1, ERBB2). 24R,25(OH)2D3 decreased proliferation in ERα66- cells (UM-SCC-11A) with the 10-7M dosage. It also increased apoptotic markers (TUNEL, P53, BAX/BCL2 expression). EMT and metastatic markers (ERBB2, SNAI1 expression) as well as bone metastatic markers (OPG/RANKL) were reduced. Conversely, in ERα66+ cells (UM-SCC-12) 24R,25(OH)2D3 had the opposite effect, increasing proliferation with 10-7M 24R,25(OH)2D3 treatment as well as decreasing DNA fragmentation (TUNEL). Treatment with 24R,25(OH)2D3 increased total P53 content and had no effect on EMT, BAX/BCL2 or other metastatic markers (SNAI1, ERBB2, OPG, RANKL). These data show that 24R,25(OH)2D3 is functionally active in laryngeal cancer tumor regulation. Contrary to results shown in breast cancer in which 24R,25(OH)2D3 was pro-tumorigenic in ERα66- cells, 24R,25(OH)2D3 was anti-tumorigenic in ERα66- laryngeal cancer cells. While this difference may be a result of a difference in other aspects of the two ERα66- cell lines, these results provide evidence of the therapeutic potential of 24R,25(OH)2D3 in laryngeal cancer and its ERα66 dependent effect. Citation Format: Cydney D. Dennis, David Joshua Cohen, Barbara D. Boyan, Zvi Schwartz. The vitamin D3 metabolite, 24R,25(OH)3D3 is differentially tumorigenic in ER+ and ER- laryngeal cancer cells in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2693.

Abstract LB526: HBI-2375, a selective inhibitor of MLL1-WDR5 interaction, possesses desirable preclinical characteristics to be pursued in IND enabling studies and future clinical investigations

Abstract 92130MLL1 (mixed lineage leukemia1) interaction with WDR5 (WD repeat domain 5) is critical for MLL1 histone methyl transferase (H3K4me) activity leading to transcriptional regulation of gene expression. MLL1 dysregulation such as its translocation at chromosome 11q23 results in generation of MLL1 fusion genes and leukemia development. Additionally, recent studies indicated a role for MLL1-WDR5 interaction in solid tumors progression via directly affecting cancer cells or by suppression of tumor immunity. Therefore, MLL1-WDR5 interaction is an important target in cancer drug discovery, both in leukemia and in solid tumors. HUYABIO recently completed acquisition of a novel series of MLL1-WDR5 inhibitors and conducted an additional SAR campaign to identify a lead compound with greater efficacy and PK (pharmacokinetics)characteristics. Characterization of a series of analogs identified HBI-2375 as a selective inhibitor of MLL1-WDR5 interaction. TR-FRET peptide binding assay indicated potent binding of HBI-2375 to WDR5 (IC50=4.48 nM) and in vitro cell potency studies showed inhibition of proliferation in MV4;11 leukemia cells by HBI-2375 (IC50=3.17 µM).Furthermore, HBI-2375 was stable in liver S9 and whole blood and displayed reasonable PK properties (Cmax, T1/2, AUC and F%) in preclinical testing in mouse and dog. In vivo efficacy studies in MV4;11 model showed 86% and 77% tumor growth inhibition in HBI-2375 treated tumors when dosed at 80 mpk or 40 mpk (po, qd x 21) respectively. In addition, HBI-2375 was found to be well tolerated since body weight data in the recipients did not show a significant change throughout the treatment. Interestingly, ex vivo tumor analysis showed a reduction in H3K4me in both 40 and 80 mpk samples, further validating target-driven activity of HBI-2375. These data resulted in the nomination of HBI-2375 as the lead clinical candidate for the current IND enabling studies and future investigations in the clinic. Citation Format: Farbod Shojaei, J E. Semple, Che Fang, Jill M. Ricono, Fairooz Kabbinavar, Bob Goodenow, Mireille Gillings. HBI-2375, a selective inhibitor of MLL1-WDR5 interaction, possesses desirable preclinical characteristics to be pursued in IND enabling studies and future clinical investigations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB526.

Tumor protein D52 (TPD52) affects cancer cell metabolism by negatively regulating AMPK.

The AMP-activated protein kinase (AMPK) is a central regulator of energy homeostasis, with deregulation leading to cancer and other diseases. However, how this pathway is dysregulated in cancer has not been well clarified. Using a tandem affinity purification/mass-spec technique and biochemical analyses, we identified tumor protein D52 (TPD52) as an AMPKα-interacting molecule. To explore the biological effects of TPD52 in cancers, we conducted biochemical and metabolic assays in vitro and in vivo with cancer cells and TPD52 transgenic mice. Finally, we assessed the clinical significance of TPD52 expression in breast cancer patients using bioinformatics techniques. TPD52, initially identified to be overexpressed in many human cancers, was found to form a stable complex with AMPK in cancer cells. TPD52 directly interacts with AMPKα and inhibits AMPKα kinase activity in vitro and in vivo. In TPD52 transgenic mice, overexpression of TPD52 leads to AMPK inhibition and multiple metabolic defects. Clinically, high TPD52 expression predicts poor survival of breast cancer patients. The findings revealed that TPD52 is a novel regulator of energy stress-induced AMPK activation and cell metabolism. These results shed new light on AMPK regulation and understanding of the etiology of cancers with TPD52 overexpression.