Non-cancerous Cells Research Articles

A novel small molecule Enpp1 inhibitor improves tumor control following radiation therapy by targeting stromal Enpp1 expression

The uniqueness in each person’s cancer cells and variation in immune infiltrates means that each tumor represents a unique problem, but therapeutic targets can be found among their shared features. Radiation therapy alters the interaction between the cancer cells and the stroma through release of innate adjuvants. The extranuclear DNA that can result from radiation damage of cells can result in production of the second messenger cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) by cyclic GMP-AMP synthase (cGAS). In turn, cGAMP can activate the innate sensor stimulator of interferon genes (STING), resulting in innate immune activation. Ectonucleotide pyrophosphatase/phosphodiesterase 1 (Enpp1) is a phosphodiesterase that can be expressed by cancer cells that can degrade cGAMP, thus can decrease or block STING activation following radiation therapy, impairing the innate immunity that is critical to support adaptive immune control of tumors. We observed that many human and murine cancer cells lack Enpp1 expression, but that Enpp1 is expressed in cells of the tumor stroma where it limits tumor control by radiation therapy. We demonstrate in preclinical models the efficacy of a novel Enpp1 inhibitor and show that this inhibitor improves tumor control by radiation even where the cancer cells lack Enpp1. This mechanism requires STING and type I interferon (IFN) receptor expression by non-cancer cells and is dependent on CD8 T cells as a final effector mechanism of tumor control. This suggests that Enpp1 inhibition may be an effective partner for radiation therapy regardless of whether cancer cells express Enpp1. This broadens the potential patient base for whom Enpp1 inhibitors can be applied to improve innate immune responses following radiation therapy.

In vitro studies of the combination between berberine chloride and a Copper(II) complex against Triple negative breast cancer

In this study, the copper(II) complex [Cu(chromoneTSC)Cl2]•0·.5H2O•0.0625C2H5OH (where chromoneTSC = (E)-N-Ethyl-2-((4-oxo-4H-chromen-3-yl)methylene)-hydrazinecarbothioamide) was synthesized and characterized; then used to carry out in vitro studies in combination with berberine chloride (BBC). The ligand and complex were characterized by elemental analysis, FTIR and NMR (1H and 13C) spectroscopy, and conductivity measurements. The cytotoxic effect was analyzed by using the CCK-8 viability assay in cancer MDA-MB-231 VIM RFP and non-cancer MCF-10A cell lines. The IC50 values for the complex and BBC were 21.2 ± 1.6 and 48.3 ± 2.4 μM, respectively at 24 h incubation, while the IC50 value of the combination treatment was 9.3 ± 1.5 in cancer cells. The co-treatment group significantly increased the number of cells in G2 phase, indicating the growth arrest of cancer cells. Moreover, the combination group showed induction of both intrinsic and extrinsic apoptotic pathways. There was also a study on the effect of the combination treatment on receptor-interacting serine/threonine-protein kinase 3 (RIPK3) and mixed lineage kinase domain-like pseudokinase (MLKL) as biomarkers of necroptosis. The results showed activation of necroptosis after treatment with the combination of the copper complex and BBC via the activation of RIPK3–MLKL pathway.

Unraveling mechanism and enhancing selectivity of a RuII-bis-bipyridyl-morphocumin complex with RAFT-generated glycopolymer exploiting Warburg effect in cancer.

Warburg effect, generates increased demand of glucose in cancer butis relatively underexplored phenomenon in existing commercial anti-cancer drugs. Here, we present aRu(II)-bis-bipyridyl-morphocumin complex (2) encapsulated in a self-assembling glucose-functionalized co-polymer P(G-EMA-co-MMA) (where G = glucose; MMA = methyl methacrylate; EMA = ethyl methacrylate), designed to exploit Warburg effect and enhance cancer selectivity. The P(G-EMA-co-MMA) polymer, synthesized via reversible-addition fragmentation chain transfer (RAFT) polymerization, has a number average molecular weight (Mn, NMR) of 8000 g/mol. Complex 2, stable in aqueous media, selectively releases a cytotoxic, lysosome-targeting compound morphocumin in the presence of excessH2O2, a reactive oxygen species (ROS) prevalent in tumor microenvironments. Complex 2 promotes ROS accumulation enhancing morphocumin release through a synergistic domino effect. Comparatively,2 outperforms its curcumin Ru(II) complex (1) in solution stability, organelle specificity, and cellular mechanisms. Both 1 and 2 exhibit phototherapeutic effects under low-intensity visible light, but their dark toxicity surpasses their photoactivity, highlighting they aresuperior as chemotherapeutic agents. Complex 2 induces apoptosis via mitochondrial pathway with 9-fold increase in selectivity for pancreatic cancer cells (MIA PaCa-2) over non-cancerous HEK293 cells when delivered encapsulated inglucose-conjugated polymer (DP@2)activityenhance further5-fold by glucose deprivation underscoring their potential in targeted cancer therapy.

Molecular Hybrids of Quinoline and Sulfonamide: Design, Synthesis and in Vitro Anticancer Studies.

Molecular hybrids of diversely functionalized quinoline and sulfonamide have been designed. Multistep synthetic strategies have been used for the synthesis. The anti-cancer properties have been evaluated against various cancer cell lines including HCT116, A549, U2OS, CCRF-CEM, Jurkat, MOLT-4, RAMOS, and K562. Non-cancer cell lines MRC-5 and BJ were also included for comparison. When examining the effects on A549, HCT116, and U2OS cells, all tested compounds exhibited limited potency with IC50 values exceeding 50 μM, indicating weak activity against these cell lines. Against the ITK high cells Viz. are Jurkat, CCRF-CEM and MOLT-4, 9 e, 9 p and 9 j found to the maximum potent compounds with IC50 values of 7.43±7.40 μM, 13.19±1.25 μM and 5.57±7.56 μM respectively. Similarly, in the BTK high cells screenings, 9 n and 9 e molecules with an IC50 value of 2.76±0.79 μM and 5.47±1.71 μM against RAMOS and K562 respectively are highly potent. Interestingly, all the molecules have exhibited IC50 value >50 μM against the non-cancer cells (MRC-5 and BJ), which indicates the promising non-cytotoxic nature of the molecules.

Self-assembled Lipid Nanoparticles for Killing Triple Negative Breast Cancer Cells.

Triple negative breast cancers (TNBCs) lacking estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) on their cell surfaces are highly aggressive, difficult-to-treat and often relapse. Herein, we report on the self-assembled lipid nanoparticles (LNPs) of two new pegylated lipopeptides for killing TNBCs (MDA-MB-231). The pegylated lipopeptides were synthesized by conjugating an n-hexadecyl hydrophobic tail to one end of a (PEG)27 unit the other distal end of which was covalently grafted with two previously reported tumor targeting RGDK- and CGKRK- peptides. The SEM images of the self-assembled LNPs formed upon dissolution of the pegylated lipopeptides in aqueous medium revealed formation of spherical aggregates. The degree of cellular uptake for the self-assembled LNPs formed by the pegylated CGKRK-lipopeptide were found to be significantly higher than that for the self-assembled LNPs formed by the pegylated RGDK-lipopeptide in MCF-7, MDA-MB-231, HEK-293 and HFF cells. Notably, about 60 % TNBCs (MDA-MB-231 cells) were killed upon treatment with commercially available potent JAK2 inhibitor (WP 1066) loaded LNPs of the pegylated RGDK-lipopeptide. Contrastingly, the same treatment killed only about 20 % non-cancerous HEK-293 cells. The self-assembled pegylated LNPs described herein open the door for undertaking preclinical studies in animal models for TNBCs.

Morphology of mitochondrial network in disseminated endometriosis cells in spontaneous pneumothorax diagnostic process.

Circulating endometrial cells (CECs) have emerged as a new biomarker of advanced disease in women with endometriosis. The identification of several subtypes of CECs (e.g., stem cell-like, epithelial, glandular, stromal) has opened the way for characterization of endometriosis-associated CECs. This study focused on the isolation and characterization of CECs and disseminated endometrial cells (DECs) in patients with spontaneous pneumothorax (SP). The primary objective was to differentiate between cancer and non-cancer cells in patients with no previous cancer diagnosis. The MetaCell® size-based separation protocol was used to enrich CECs/DECs. Evaluation of the captured cells by 3D microscopy was performed using a NANOLIVE™ microscope using a holographic approach. Based on gene expression analysis (GEA), we can conclude that mitochondria are much more active in primary tumors compared to endometriosis tissue (e.g. MT-ND1, MT-ATP6 genes). The culture of DECs is made of stromal, stem and immune cells. In vitro culture of DECs is characterized by an increase in the epithelial marker KRT18. Similarly, NFE2L2, a proerythroid factor, is also elevated. Further, a significant decrease in the amount of stem and immune cells was observed in the cell culture of DECs. The data presented here show how morphologically plastic the changes in the mitochondrial network can be and how cells can reflect them at the level of gene expression. The markers identified could help in the accompanying diagnostic process of the spontaneous pneumothorax in women of reproductive age.

In Vitro Anticancer Effects of Aqueous Leaf Extract from Nepeta nuda L. ssp. nuda

Despite significant efforts, cancer remains the second leading cause of mortality worldwide. The medicinal plant Nepeta nuda L. represents a valuable source of biologically active compounds with pharmacological activities including antioxidant, anti-inflammatory, antimicrobial, and antiviral. This study aimed to assess the antiproliferative potential and mechanisms of action of aqueous extract from the leaves of wild-grown N. nuda. Cancer cell lines, MDA-MB-231, MCF7 (breast), HT29, Colon 26 (colon), and HepG2 (liver cancer), and a non-cancerous skin cell line, BJ, were assessed for antiproliferative activity by MTT assay and observation of cell morphological alterations. The cancer cell line that was most sensitive to the extract was further studied for apoptotic alterations by Annexin V/propidium iodide staining, colony-forming assay, and qRT-PCR analysis. The results revealed that the plant extract inhibited the proliferation of all investigated cancer cell lines with the strongest cytostatic effect on Colon 26 cells with a half maximal inhibitory concentration (IC50) value of 380.2 μg/mL and a selectivity index (SI) of 3.5. The extract significantly inhibited the ability of cells to form colonies, exhibited considerable proapoptotic potential involving the participation of the CASP8 gene, and increased the expression levels of ATG3 and the BECN1 gene, which suggests a role of autophagic cell death in the antitumor action.

Biological Evaluation of a Rhodium(III) Bipyridylsulfonamide Complex: Effects on Mitochondrial Dynamics and Cytoskeletal Remodeling in Breast Cancer Cells.

Rhodium(III) complexes have gained attention for their anticancer potential. In this study, we investigated a rhodium(III) bipyridylsulfonamide complex (2) and its ligand (L) for their effects on breast cancer (SKBr3) and noncancerous mammary cells (HB2). Both compounds significantly reduced oxidative phosphorylation (OXPHOS) and mitochondrial function in SKBr3 cells while sparing HB2 cells. Compound 2 also increased glycolysis in both lines, suggesting a metabolic shift. Mitochondrial size and shape were altered, particularly in SKBr3 cells. Additionally, both compounds reduced cancer cell migration by disrupting actin cytoskeleton organization and the Rac1/VASP signaling pathway. These findings suggest that the rhodium(III) bipyridylsulfonamide complex selectively impairs mitochondrial dynamics and cell migration in cancer cells while sparing healthy cells, providing insight into its mechanism of action and toward its use as targeted anticancer therapy. This study lays the groundwork for future in vivo studies and further optimization of these metal-based therapeutics for clinical applications.

POCOP-Ni(II) pincer compounds derived from phloroglucinol. Cytotoxic and antioxidant evaluation.

POCOP-Ni(II) pincer compounds have primarily been explored as catalysts, but their potential biological activity has been scarcely studied. To address this gap, we evaluated the anticancer and antioxidant potential of four POCOP-Ni(II) complexes derived from phloroglucinol. A comprehensive supramolecular analysis, based on single-crystal X-ray diffraction (DRX) structures, was conducted using Hirshfeld surfaces and non-covalent interaction analysis. The cytotoxicity of all complexes was systematically assessed against various cancerous cell lines, as well as a non-cancerous cell line (COS-7). The results revealed that complexes 1b and 1c exhibited remarkable antiproliferative activity, with IC50 values ranging from 2.43 to 7.85μM against cancerous cell lines U251, K562, HCT-15, MCF-7, and SK-LU-1. To further elucidate their mechanism of action, a competitive fluorescence displacement assay with ethidium bromide (EB) suggested that these complexes possess the ability to intercalate with DNA. This multifaceted investigation not only enhances our understanding of the biological potential of POCOP-Ni complexes but also provides valuable insights into their structural features and interactions, paving the way for future exploration in both catalytic and therapeutic domains.

Pioneering Advances and Innovative Applications of Mesoporous Carriers for Mitochondria-Targeted Therapeutics.

Mitochondria, known as the cell's powerhouse, play a critical role in energy production, cellular maintenance, and stemness regulation in non-cancerous cells. Despite their importance, using drug delivery systems to target the mitochondria presents significant challenges due to several barriers, including cellular uptake limitations, enzymatic degradation, and the mitochondrial membranes themselves. Additionally, barriers in the organs to be targetted, along with extracellular barriers formed by physiological processes such as the reticuloendothelial system, contribute to the rapid elimination of nanoparticles designed for mitochondrial-based drug delivery. Overcoming these challenges has led to the development of various strategies, such as molecular targeting using cell-penetrating peptides, genomic editing, and nanoparticle-based systems, including porous carriers, liposomes, micelles, and Mito-Porters. Porous carriers stand out as particularly promising candidates as drug delivery systems for targeting the mitochondria due to their large pore size, surface area, and ease of functionalisation. Depending on the pore size, they can be classified as micro-, meso-, or macroporous and are either ordered or non-ordered based on both size and pore uniformity. Several methods are employed to target the mitochondria using porous carriers, such as surface modifications with polyethylene glycol (PEG), incorporation of targeting ligands like triphenylphosphonium, and capping the pores with gold nanoparticles or chitosan to enable controlled and triggered drug delivery. Photodynamic therapy is another approach, where drug-loaded porous carriers generate reactive oxygen species (ROS) to enhance mitochondrial targeting. Further advancements have been made in the form of functionalised porous silica and carbon nanoparticles, which have demonstrated potential for effective drug delivery to mitochondria. This review highlights the various approaches that utilise porous carriers, specifically focusing on silica-based systems, as efficient vehicles for targeting mitochondria, paving the way for improved drug delivery strategies in mitochondrial therapies.

Abstract C033: Identifying and targeting prostate cancer therapy resistance mechanisms driven by the tumor microenvironment

Abstract Background: Prostate cancer is a prevalent malignancy in men, with treatment resistance posing a significant challenge to successful cure. Acquired Resistance to Therapy (ART) arises from two main components: (1) Tumor Microenvironment (TME) and (2) mutations or epigenetic changes at gene level within the tumor cells themselves. While most cancer therapies target the intrinsic biology of cancer cells, growing evidence indicates that these treatments can also harm the stromal cells within TME, potentially driving ART. The biology of ART may vary based on the types of treatments, cell types, and organ locations. Thus, this study aims to dissect ART components driven by TME-mediated cellular processes in a comprehensive manner. Hypothesis: We propose that cancer therapies induce stress responses in non-cancerous cells within the TME. These stress responses involve the secretion of cytokines, mediators, and growth factors, which we term Stress Response Secretory Programs (SRSPs). We further hypothesize that targeting SRSPs could overcome ART that helps to improve treatment specificity and efficacy. Materials and Methods: We employed comprehensive single-cell profiling of common metastatic sites included liver, prostate, lung, bone, lymph node and spleen in preclinical models to identify secreted proteins and dysregulated pathways caused by genotoxic therapies. In vitro, we evaluated treatment resistance in murine prostate epithelial cell lines (Myc-CAP) by exposing them to conditioned media (CM) from irradiated murine fibroblasts (F4M2), using cell growth assays. Results: Single-cell RNA sequencing of organ samples revealed significant expressions of SRSPs and increased inflammatory responses in liver and prostate fibroblasts as well as other stromal cells treated with carboplatin. In vitro, CM from irradiated F4M2 cells enhanced cell proliferation and survival of Myc-CAP cells in response to docetaxel treatment. Conclusion: Our findings highlight the crucial role of SRSPs in mediating ART through interactions within the TME, which alter tumor cell phenotypes. Targeting these SRSPs holds potential for overcoming treatment resistance and improving therapeutic outcomes, underscoring the need to consider TME complexity in cancer treatment strategies. Citation Format: Tony Lok Heng Chu, Armand Bankhead, Ilsa Coleman, Sander Frank, Peter S. Nelson, Tarana Arman. Identifying and targeting prostate cancer therapy resistance mechanisms driven by the tumor microenvironment [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C033.

Synthesis, characterization, and co-delivery of doxorubicin and melittin-loaded NiyMg1-yFe2O4 nanoparticles for combination therapy in MCF-7 and NIH-3T3 cell lines

The present investigation reporting about the synthesis of NiyMg1-yFe2O4 (y = 0.3, 0.5, and 0.7) nanoparticles employing the co-precipitation method. TG-DT analysis revealed that NiyMg1-yFe2O4 nanoparticles were thermally stable above 386 °C. Structural, morphological, and magnetic studies were carried out through XRD, FTIR, FESEM, HRTEM, and VSM tools. XRD analysis confirmed that the nanoparticles possessed a single-phase cubic spinel structure and the Debye-Scherrer equation employed notices a linear increase in average crystallite size with increases in both nickel concentration and annealing temperatures. Two prominent absorption peaks between 423 and 556 cm−1, observed in FTIR study corresponding to the stretching vibration of octahedral (B) and tetrahedral (A) sited characteristic of spinel ferrite. The surface morphology of the NiyMg1-yFe2O4 nanoparticles annealed at 600 °C was characterized through FESEM and HRTEM tools. FESEM micrographs emphasized a spherical shape with less agglomeration, while EDX spectra confirmed the presence of Fe, Ni, Mg, and O in the nanoparticles advocating its purity. HRTEM images further confirmed the spherical morphology of the nickel magnesium ferrite nanoparticles. Magnetic properties were measured using VSM technique, suggesting the saturation magnetization and coercivity values increased with increased nickel concentrations, showing its soft ferrimagnetic behavior and suggesting potential applications in drug delivery and storage devices. The in vitro investigation was carried out using MTT assay to ascertain the amount of cytotoxicity present in breast cancer (MCF-7) and non-cancerous (NIH-3T3) cells taken for study with help of prepared nanoparticles administered with drugs doxorubicin (DOX) and melittin (MLT).

Abstract B018: miRNA-130a promotes Breast cancer progression by inhibiting multiple tumour suppressor genes as a Gene-TF-miRNA network

Abstract Cancer is a disease that has implications worldwide, including in developing countries. It is known for its notorious ways of adapting and gaining resistance to the therapies available. Breast cancer (BC) majorly affects women and leading cause of death in women worldwide. Despite studying BC for many decades, we are not yet certain about a definitive therapy that can cure it with confidence. One of the challenges associated with treating BC is its metastasis and resistance to available therapies. Non-cancerous cells exhibit a tightly regulated transcription profile. An altered transcription profile is considered one of the main causes of the origin of cancers and their progression, including resistance and metastasis. Tumor suppressor genes are associated with mitigating any oncogenic signals induced by external factors or internal dysregulation in oncogenic genes. Changes in the expression profiles of tumour suppressor and oncogenic genes are associated with BC progression and metastasis. Transcription factors (TFs) play a significant role in marinating the tight regulation of transcription. In this study, we have analyzed Genes(N=1057), TFs(N =233) and miRNAs (N = 246) that are promoting BC from literature and publicly available databases; we have analyzed the regulation of Gene-Gene, miRNA-miRNA, TF-TF, miRNA-Gene, miRNA-TF, TF-Gene, TF-miRNA as a network of three node Feed Forward Loops (FFLs). Upon constructing 3, 4,5 and 6 node FFLs, we have identified that the miRNA130a is promoting BC by inhibiting crucial Tumor suppressor genes like TGFBR1, TGFBR2, SOD2, PTEN, SMAD4, RUNX3 etc. Citation Format: Prasanna Kumar Reddy Gayam, Jesil Mathew A, Fayaz SM. miRNA-130a promotes Breast cancer progression by inhibiting multiple tumour suppressor genes as a Gene-TF-miRNA network [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr B018.

Uncovering the Potential ofO-Prenylated 3-aryl-benzopyran Derivatives as Osteogenic and Cancer Cell Growth Inhibitory Agents.

Naturally, O-prenylation of 3-aryl-benzopyrans enhances the biological activities of these compounds. In this study, substituted O-prenylated 3-aryl-benzopyrans (21a-c, 22a-c, 23a-c, 24a-c 25a-c, 27 and 28) were synthesized and evaluated for osteogenic and cancer cell growth inhibitory potentials using cell-based in-vitro models. Amongst the target compounds, 21a, 22b, 23c, and 24c showed good osteogenic activity at 1 pM concentration, whereas 26 and 27 showed osteogenic activity at 100pM and 10nM, respectively. Compounds 21a, 22b, and 23c showed good cancer cell growth inhibitory activity against breast cancer cells (MCF-7 and MBA-231). Amongst active compounds, 27 presented the best anticancer activity against MDAMB-231 cells with selectivity towards non-cancerous cells [IC50 3.76 µM with SI 13.3]. The in-silico study of compounds showed their structural complementarities with the LBD of estrogen receptors and compliance with dragability parameters.

Drug prioritization identifies panobinostat as a tailored treatment element for patients with metastatic hepatoblastoma

BackgroundPatients with metastatic hepatoblastoma are treated with severely toxic first-line chemotherapies in combination with surgery. Yet, inadequate response of lung metastases to neo-adjuvant chemotherapy still compromises patient outcomes making new treatment strategies, tailored to more efficient lung clearance, mandatory.MethodsWe harnessed a comprehensive patient-derived xenograft platform and a variety of in vitro and in vivo assays to establish the preclinical and biological rationale for a new drug for patients with metastatic hepatoblastoma.ResultsThe testing of a library of established drugs on patient-derived xenografts identified histone deacetylase inhibitors, most notably panobinostat, to be highly efficacious on hepatoblastoma cells, as compared to non-cancerous cells. Molecularly, the anti-tumor effect of panobinostat is mediated by posttranslational obstruction of the MYC oncoprotein as a result of dual specificity phosphatase 1 upregulation, thereby leading to growth inhibition and programmed cell death. Of clinical importance, upregulation of the MYC target gene nucleophosmin 1 is indicative of response to panobinostat and associated with metastatic disease in patients with hepatoblastoma. The combination of panobinostat with the current SIOPEL 4 induction protocol, consisting of cisplatin and doxorubicin, revealed high synergies already at low nanomolar levels. The simulation of a clinical trial, with this combination therapy, in patient-derived xenograft models, and ultimately heterotypic lung metastasis mimics clearly underscored the potency of this approach.ConclusionIntegrated studies define MYC inhibition by panobinostat as a novel treatment element to be introduced into the therapeutic strategy for patients with metastatic hepatoblastoma.

DDDR-13. OPTIMIZING BRAIN CANCER THERAPY: BALANCING TUMOR ERADICATION AND NORMAL TISSUE PRESERVATION WITH BPM31510

Abstract The challenges translating glioblastoma therapies lie in safeguarding normal neuronal and glial tissue. Considering that the central nervous system has the lowest regenerative capacity, the exclusive focus on tumor eradication while neglecting normal tissue response, impedes proper healing. This oversight leads to toxicities ultimately culminating in patient demise with marginal gain in survival. BPM31510 is a nanodispersion encapsulating highly hydrophobic oxidized Coenzyme Q10 (CoQ10), allowing for delivery of supraphysiological CoQ10 concentrations into cancer cells. Initial investigations demonstrated differential responsiveness between cancer and non-cancer cells in vitro, an effect associated with high superoxide levels exclusively in cancer cells. To evaluate this divergence between cancer and non-cancer cells, a competitive co-culture cell-based assay was developed with labeled pairs of glioblastoma and non-cancer cells incubated up to three weeks with various concentrations of BPM31510 to ascertain optimal concentration leading to preferential growth of non-cancer cells. Human astrocyte (HA), mouse fibroblast (NIH 3T3), and rat astrocyte (TNC1) cells were paired with U251 human glioma, mouse GL261 and rat C6 glioma, respectively. Cell growth and spatial extent of the plate containing non-cancer or tumor cells were monitored every other day. Using doxorubicin as control, no dosage was discerned in which non-cancer cells exhibited advantage over tumor cells (i.e., either cultures became all tumor cells or both cell lines were eradicated). Using various concentrations of BPM31510, we identified an optimal dose in which cancer cells diminished while non-cancer cells predominated, reflected by markedly elevated mitochondrial superoxide levels in cancer cells prior to cell death. A co-culture cell-based assay was developed to identify an optimal dose of BPM31510 with selective preservation of non-cancer cells alongside tumor cell death. This approach elucidates developing a “fine tuning” strategy, wherein BPM31510 is titrated over time, thereby promoting normal tissue and healing, and ultimately leading to superior and prolonged functional outcomes.

DNAR-09. GB13 IS A POTENT NEOADJUVANT TREATMENT OPTION FOR IL13RA2-EXPRESSING GBM AND IMPROVES STANDARD-OF-CARE THERAPY

Abstract Glioblastoma (GBM) is the most common primary adult brain cancer and is uniformly fatal. Despite maximum safe surgical resection followed by radiation and temozolomide chemotherapy, current median survival is 14-18 months. GB13 is a novel therapy for treatment of GBM and received Orphan Drug Designation by the Food and Drug Administration (FDA) for treatment of malignant gliomas. GB13 specifically targets the tumor-restricted IL13Ra2 receptor that is highly expressed in GBM, among other cancers, but not on non-cancerous cells. IL13Ra2 represents a clinically viable target for directed therapies and is being explored by numerous approaches. Previous data demonstrate that GB13 potently kills GBM and diffuse midline glioma (DMG) cells and tumors, IL13Ra2 correlates to GB13 sensitivity and, GB13 can significantly improve the cytotoxic effects of ionizing radiation (IR) when tested in models of pediatric diffuse midline glioma. In the current studies, the ability of GB13 to enhance the effects of IR were explored using IL13Ra2-positive and negative GBM models. Major findings demonstrate that neoadjuvant GB13 treatment increases cytotoxic effects of IR, enhances cell apoptosis and decreases cell viability. These results were not observed in IL13Ra2-negative settings, demonstrating the necessity for this clinical target. Mechanistically, increased cell death was caused by sustained apoptotic signaling through caspase-mediated pathways. Preclinical models support the use of GB13 prior to radiation for decreased tumor proliferation. Using multiple models, our results identify a temporal benefit for treating GBM tumors prior to IR and this should be tested in future clinical studies.

Enhancing Phototoxicity in BODIPY-Perylene Charge Transfer Dyads by Combined Iodination and Mesylation.

The uptake and phototoxicity of a family of BODIPY-perylene charge transfer dyads are compared in live cancer and non-cancer cell lines to evaluate their performance in imaging and photodynamic therapy (PDT). The impact of iodination and mesylation of the meso position of the compounds on their optical properties, cell uptake and toxicity are compared. Notably, across all derivatives the probes were minimally dark toxic up to 50 μM, (the maximum concentration tested), but exhibited outstanding phototoxicity with nanomolar IC50 values and impressive phototoxic indices (PI, ratio of dark IC50 to light IC50), with best performance for the mesylated iodinated derivative MB2PI, which had a PI of >218 and >8.9 in MCF-7 cells and tumour spheroids respectively. This is significantly higher than non-iodinated analogue MB2P in MCF-7 cells with an observed PI of >109 and slightly higher than MB2PI in spheroids with a PI of >8. This compound also showed interesting emission spectral variation with localisation that responded to stimulation of inflammation. Additional studies confirmed efficient singlet oxygen generation by the BODIPYs, suggesting a Type II mechanism of phototoxicity. Overall, the data indicates that combining charge transfer and iodination is an effective strategy for enhancing phototherapeutic capacity of BODIPY PS.

DeSide: A unified deep learning approach for cellular deconvolution of tumor microenvironment

Cellular deconvolution via bulk RNA sequencing (RNA-seq) presents a cost-effective and efficient alternative to experimental methods such as flow cytometry and single-cell RNA-seq (scRNA-seq) for analyzing the complex cellular composition of tumor microenvironments. Despite challenges due to heterogeneity within and among tumors, our innovative deep learning-based approach, DeSide, shows exceptional accuracy in estimating the proportions of 16 distinct cell types and subtypes within solid tumors. DeSide integrates biological pathways and assesses noncancerous cell types first, effectively sidestepping the issue of highly variable gene expression profiles (GEPs) associated with cancer cells. By leveraging scRNA-seq data from six cancer types and 185 cancer cell lines across 22 cancer types as references, our method introduces distinctive sampling and filtering techniques to generate a high-quality training set that closely replicates real tumor GEPs, based on The Cancer Genome Atlas (TCGA) bulk RNA-seq data. With this model and high-quality training set, DeSide outperforms existing methods in estimating tumor purity and the proportions of noncancerous cells within solid tumors. Our model precisely predicts cellular compositions across 19 cancer types from TCGA and proves its effectiveness with multiple additional external datasets. Crucially, DeSide enables the identification and analysis of combinatorial cell type pairs, facilitating the stratification of cancer patients into prognostically significant groups. This approach not only provides deeper insights into the dynamics of tumor biology but also highlights potential therapeutic targets by underscoring the importance of specific cell type or subtype interactions.

Synthesis and Biological Evaluation of N-(1H-Indol-6-ylmethyl)benzenesulfonamide Analogs as Metabolic Inhibitors of Mitochondrial ATP Production in Pancreatic Cancer Cells.

A library of 26 indolyl sulfonamides and 12 amide and ester analogs based upon the 6-indolyl framework has been synthesized in an effort to target pancreatic cancer. The cytotoxicity of the indolyl sulfonamide compounds has been determined using a traditional (48-h compound exposure) assay against 7 pancreatic cancer cell lines and 1 non-cancerous cell line. The potential role of the compounds as metabolic inhibitors of ATP production was evaluated using a rapid screening (2-h compound exposure) assay developed within our laboratories. The IC50 values of the active compounds were determined using the rapid assay and six compounds displayed an IC50 value <5 μM against one or more pancreatic cancer cell lines. The ester analogs also display activity as potential metabolic inhibitors of ATP production with four of the six compounds displaying an IC50 value <5 μM against one or more pancreatic cancer cell lines.