Reduced Tumor Development Research Articles

Inhibition of mitochondrial OMA1 ameliorates osteosarcoma tumorigenesis

OMA1 is an ATP-independent zinc metalloprotease essential for maintaining mitochondrial homeostasis and plays a vital role in tumorigenesis. Depending on the type of cancer, a decrease in OMA1 expression has been linked to a varying prognosis for patients. The role of OMA1 in human osteosarcoma (OS), one of the most prevalent malignant bone tumors, remains elusive. Here, we observed elevated OMA1 expression in OS tumor tissues from four patients with advanced OS. Knockout of OMA1 in OS cells significantly reduces OS tumor weight and size, and lung metastatic nodules in BALB/c nude mice. Immunohistochemistry analysis showed a significant decrease in Ki67 and an increase in Cleaved-caspase 3 in OMA1 knockout tumor samples. Mechanistically, we found that OMA1 deficiency increases the levels of PINK1 and Parkin and consequently induces excessive mitophagy, leading to increased apoptosis and reduced cell proliferation and invasion in OS cells. Specifically, OMA1 deficiency reduces the amount of cytosolic p53 and p53-associated cytosolic Parkin but increases mitochondrial p53, which may lead to enhanced apoptosis. Regarding the effect on cell proliferation and invasion, loss of OMA1 reduces mitochondrial ROS levels and increases cytosolic glycogen synthase kinase 3β (GSK3β) levels, thereby increasing interaction between GSK3β and β-catenin and then reducing cytosolic and nuclear β-catenin. This contributes to reduced cell proliferation and migration in OMA1-deficient cells. Moreover, we found that ciclopirox (CPX), an antifungal drug, induces OMA1 self-cleavage and L-OMA1 degradation in cultured OS cells. CPX also reduces tumor development of control OS cells but not OMA1-deficient OS cells in mice. These findings strongly support the important role of OMA1 in OS tumorigenesis and suggest that OMA1 may be a valuable prognostic marker and a promising therapeutic target for OS.

Heparin-binding EGF-like growth factor via miR-126 controls tumor formation/growth and the proteolytic niche in murine models of colorectal and colitis-associated cancers

MicroRNAs, including the tumor-suppressor miR-126 and the oncogene miR-221, regulate tumor formation and growth in colitis-associated cancer (CAC) and colorectal cancer (CRC). This study explores the impact of the epithelial cytokine heparin-binding epidermal growth factor (HB-EGF) and its receptor epidermal growth factor receptor (EGFR) on the pathogenesis of CAC and CRC, particularly in the regulation of microRNA-driven tumor growth and protease expression. In murine models of CRC and CAC, lack of miR-126 and elevated miR-221 expression in colonic tissues enhanced tumor formation and growth. MiR-126 downregulation in colon cells established a pro-tumorigenic proteolytic niche by targeting HB-EGF-active metalloproteinase-7, -9 (MMP7/MMP9), disintegrin, and metalloproteinase domain-containing protein 9, and modulating chemokine-mediated recruitment of HB-EGF-loaded inflammatory cells. Mechanistically, downregulation of HB-EGF and EGFR in the colon suppressed miR-221 and enhanced miR-126 expression via activating enhancer-binding protein 2 alpha. Reintroducing miR-126 reduced tumor development and HB-EGF expression. Combining miR-126 reintroduction, which targets specific HB-EGF-active proteases but not ADAM17, with MMP inhibitors like Batimastat or Marimastat effectively suppressed tumor growth. This combination normalized protease expression and balanced miR-126 and miR-221 levels in developing and growing tumors. These findings demonstrate that suppressing HB-EGF and EGFR1 shifts the balance from oncogenic miR-221 to tumor-suppressive miR-126 action. Consequently, normalizing miR-126 expression could open new avenues for treating patients with CAC and CRC, and this normalization is intertwined with the anticancer efficacy of MMP inhibitors.

RBBP4: A novel diagnostic and prognostic biomarker for non-small-cell lung cancer correlated with autophagic cell death.

Non-small-cell lung cancer (NSCLC) often presents at later stages, typically associated with poor prognosis. Autophagy genes play a role in the progression of tumors. This study investigated the clinical relevance, prognostic value, and biological significance of RBBP4 in NSCLC. We assessed RBBP4 expression using the GSE30219 and TCGA NSCLC datasets and NSCLC cells, exploring its links with clinical outcomes, tumor immunity, and autophagy genes through bioinformatics analysis after transcriptome sequencing of RBBP4-knockdown and control PC9 cells. We identified differentially expressed genes (DEGs) and conducted Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway enrichment, and protein-protein interaction network analyses. The significance of autophagy-related DEGs was evaluated for diagnosis and prognosis using the GSE30219 dataset. Experiments both invivo and invitro explored the biological mechanisms behind RBBP4-mediated autophagic cell death in NSCLC. RBBP4 overexpression in NSCLC correlates with a poorer prognosis. Eighteen types of immune cell were significantly enriched in cultures that had low RBBP4 expression compared high expression. DEGs associated with RBBP4 are enriched in autophagy pathways. Transcriptomic profiling of the PC9 cell line identified autophagy-related DEGs associated with RBBP4 that exhibited differential expression in NSCLC, suggesting prognostic applications. Invitro experiments demonstrated that RBBP4 knockdown induced autophagy and apoptosis in PC9 cells, promoting cell death, which was inhibited by 3-MA. Invivo, targeted siRNA against RBBP4 significantly reduced tumor development in PC9 cell-injected nude mice, elevating autophagy-related protein levels and inducing apoptosis and necrosis in tumor tissues. In NSCLC, RBBP4 upregulation correlates with poor prognosis and altered immunity. Its knockdown induces autophagic cell death in NSCLC cells. These results indicate RBBP4 as a potential NSCLC diagnostic marker and its autophagy modulation as a prospective therapeutic target.

Organoruthenium metallocycle induced mutation in gld-1 tumor suppression gene in JK1466 strain and appreciable lifespan expansion

Four Ru(II) complexes (A2-A5) were synthesized from the reaction of coumarin Schiff base ligands (7da2-tsc, 7da3-mtsc, 7da4-etsc and 7da5-ptsc) with [RuHCl(CO)(PPh3)3]. The compounds were characterized by FT-IR, UV–Vis, 1H, 13C and 31P NMR, mass spectrometry and crystallographic analysis. Calf Thymus DNA (CT-DNA) binding studies revealed the intercalative mode of binding of the complexes with DNA. The results of Bovine serum albumin (BSA) binding studies established the interaction between BSA followed static quenching mechanism. The cytotoxic effects of the complexes and the ligands were evaluated against breast (MCF-7 and MDA-MB-231) and lung carcinoma cell lines (A549 and NCI-H460) using MTT assay. Complex A4 demonstrated potent cytotoxic effects on both breast and lung cancer cells. Furthermore, morphological observations and FACS analysis showed the decrease in cell density by complex A4 by induced morphological changes and apoptotic body formation and cell death in both breast and lung cancer cells. Moreover, the invertebrate model Caenorhabditis elegans was employed to assess the in vivo anticancer activity of compound A4. The findings indicated that the treatment with A4 reduced tumor development and significantly extended organismal lifespan by 64 % in the tumoral strain JK1466 without adversely affecting essential physiological functions of the worm. Additionally, A4 demonstrated an upregulation of two crucial antioxidant defense genes. Overall, these results suggested that the compound A4 can be a potential candidate with novel chemotherapeutic applications.

Macrophages, IL-10, and nitric oxide increase, induced by hyperglycemic conditions, impact the development of murine melanoma B16F10-Nex2

Epidemiological studies show a strong correlation between diabetes and the increased risk of developing different cancers, including melanoma. In the present study, we investigated the impact of a streptozotocin (STZ)-induced hyperglycemic environment on B16F10-Nex2 murine melanoma development. Hyperglycemic male C57Bl/6 mice showed increased subcutaneous tumor development, partially inhibited by metformin. Tumors showed increased infiltrating macrophages, and augmented IL-10 and nitric oxide (NO) concentrations. In vivo neutralization of IL-10, NO synthase inhibition, and depletion of macrophages reduced tumor development. STZ-treated TLR4 KO animals showed delayed tumor development; the transfer of hyperglycemic C57Bl/6 macrophages to TLR4 KO reversed this effect. Increased concentrations of IL-10 present in tumor homogenates of hyperglycemic mice induced a higher number of pre-angiogenic structures in vitro, and B16F10-Nex2 cells incubated with different glucose concentrations in vitro produced increased levels of IL-10. In summary, our findings show that a hyperglycemic environment stimulates murine melanoma B16F10-Nex2 primary tumor growth, and this effect is dependent on tumor cell stimulation, increased numbers of macrophages, and augmented IL-10 and NO concentrations. These findings show the involvement of tumor cells and other components of the tumor microenvironment in the development of subcutaneous melanoma under hyperglycemic conditions, defining novel targets for melanoma control in diabetic patients.

Abstract 5326: Selective deletion of estrogen receptor β in lung epithelium suppresses K-ras mutant lung tumorigenesis

Abstract Although lung cancer is the most prolific cause of global cancer-related deaths, outcomes by sex have gained traction as a means to stratify patients and tailor therapies accordingly. In recent years, females have begun to shoulder a greater burden of lung cancer diagnoses, specifically within the lung adenocarcinoma (LUAD) subtype; conversely, mortality is worsened for males. However, other sex-dependent effects in lung cancer are harder to predict. For example, hormone replacement therapy for post-menopausal women has been shown to predict better survival in some cohorts while auguring worse outcomes in others. Similar controversy exists regarding menopausal status, gender affirming care, and castration/oophorectomy status. However, there is consensus that these differential effects relate to a patient’s sex hormone composition. Estrogen, a sex hormone abounding in females, boasts well-described pro-tumor signaling in breast cancer. Unfortunately, its role in lung cancer remains elusive. Estrogen is reported to exert inflammatory properties in the lung environment, to drive immune function, and to induce proliferation of lung cancer cells. This functional antagonism has led to questions about how these compartment-specific effects are balanced. In the lung, estrogen signaling occurs mainly via estrogen receptor beta (ERβ), a nuclear hormone receptor highly expressed in lung tissue and lung tumors. ERβ is understudied relative to its sibling ERα, a well-known oncogene in breast cancer. While it is tempting to analogize ERα oncogene function to ERβ, the literature lacks consensus on the role of ERβ in lung cancer, with some authors classifying it as an oncogene and others as a tumor suppressor. In LUAD, Kras mutations encompass a quarter of driver mutations, and interaction of Kras with ERβ is understudied. Here, we tested the effects of genetic ablation of ERβ in a murine model of Kras mutant LUAD (KM-LUAD) termed CCSPCre/LSL-KrasG12D (CC-LR). Esr2 floxed mice were crossed with CC-LR mice to create a lung epithelial-specific conditional knockout of ERβ (LR/Esr2Δ/Δ). LR/Esr2Δ/Δ mice show significant decrease in tumor burden and total lung immune infiltration, indicating reduced tumor development and tumor-promoting inflammation. Of note, this effect manifests regardless of sex. RNA and protein-level analysis further suggest that ERβ deletion alters the balance of inflammatory transcription factors, resulting in skewing from STAT3- to NF-κB-driven inflammation and a net anti-tumor response. These data suggest that ERβ offers a promising early target in nascent KM-LUAD tumors with therapeutic intervention possible regardless of sex-specific effects. Acknowledgement: supported by NIH/NCI grant awarded to Seyed Javad Moghaddam (R01 CA225977); Michael J Clowers partly supported by NIH/NCI (F31 CA261121); Annamarie L Thompson supported by CPRIT Research Training Award (RP210028). Citation Format: Michael Joseph Clowers, Annamarie L. Thompson, Carlos Ignacio Rodriguez Reyna, Shanshan Deng, Seyed Javad Moghaddam. Selective deletion of estrogen receptor β in lung epithelium suppresses K-ras mutant lung tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5326.

Abstract 4652: Combined inhibition of MCL-1 and MEK reduces tumor growth in a triple-negative/inflammatory breast cancer, MEK-resistant xenograft mouse model

Abstract Background: Triple-negative breast cancer (TNBC) and inflammatory breast cancer (IBC) are highly aggressive subtypes of invasive breast cancer. Our previous work showed that the MEK inhibitor (MEKi) selumetinib (AZD6244) prevents lung metastasis in a xenograft model. However, clinical studies have shown that MEKi’s as single agents have only modest activity against solid tumors. Using a genome-wide synthetic lethal siRNA screening, we identified myeloid cell leukemia-1 (MCL1) as a potential contributor to selumetinib resistance. MCL-1 is an anti-apoptotic protein associated with cell immortalization, transformation, and chemoresistance in numerous cancers. We hypothesized that MCL-1 promotes MEKi resistance in TNBC/IBC and inhibition of MCL-1 in the presence of AZD6244 overcomes this MEKi resistance. Methods & Results: To understand MCL1’s role in MEKi resistance, we established two AZD6244 -resistant TNBC/IBC cell lines: MDA-MB-231-R and SUM149-R. These cells showed increased cell viability, colony formation, migration, cells in G1 phase, CD44+CD24low, mammosphere count, anchorage-independent growth and high MCL-1 expression. MCL-1’s biological role was evaluated by transient and stably knock down (KD) of MCL-1 in the resistant cells which showed decreased cell viability, colony formation, and increased apoptosis. To understand the mechanism mediating apoptosis, we examined the expression of MCL-1’s binding partners in the MCL-1-KD-R cells in the presence of AZD6244. These cells showed a high expression of the PUMA, NOXA, BAK, and BAX in the presence of AZD6244. Further, we treated resistant cells with MEKi AZD6244 and an MCL-1i (AZD5991) that confirmed restoration of MEK sensitivity in both resistant cell lines, resulting in decreased cell viability, colony formation and increased apoptosis. We compared tumor formation capability between SUM149 parental and SUM149 resistant cells by injecting them in nu/nu mice and observed a time-dependent increase (avg. 4-fold) in tumor growth with SUM149-R cells compared to parental cells (P≤0.0001). Moreover, IHC analysis showed high expression of MCL-1 (P≤0.01), Ki67 (ns), CD44 (P≤0.0001), and ALDHA1 (P≤0.0001) in the mammary gland tumor tissues of SUM149-R-injected mice compared to SUM149-P-injected mice. Next, we evaluated the therapeutic effect of AZD6244 in mice harboring SUM149-R - MCL-1-deficient tumors. AZD6244 treatment significantly reduced tumor development in mice harboring SUM149-R - MCL-1-KD tumors compared to control mice (P<0.0001), suggesting that MCL-1 mediates MEK resistance in triple-negative/inflammatory breast cancer. Conclusion: Our results suggest that MCL-1 is important for inducing acquired MEK resistance and combining an MCL-1i with a MEKi could improve treatment outcomes of patients with TNBC or IBC who have developed resistance to single-agent MEKi. Citation Format: Mohd Mughees, Alex Tan, Lakesla Iles, Michael White, Debu Tripathy, Savitri Krishnamurthy, Jian Wang, Wendy A. Woodward, Geoffrey Bartholomeusz, Chandra Bartholomeusz. Combined inhibition of MCL-1 and MEK reduces tumor growth in a triple-negative/inflammatory breast cancer, MEK-resistant xenograft mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4652.

Abstract 6675: Dissecting the relationship between mucin 5AC and the lung/gut microbiome in the pathogenesis of K-ras mutant lung cancer

Abstract Non-small cell lung cancer, including its most common subtype, lung adenocarcinoma (LUAD), is one of the most prevalent and lethal forms of cancer worldwide. This is largely due to patients having progressed to an advanced stage of disease by the time of diagnosis. Thus, better understanding the pathogenesis of early LUAD is paramount for the identification of key targets and development of preventive and therapeutic strategies for clinical management of the disease. KRAS mutations are the most common oncogenic driver alteration in human LUAD. Previously, we have found that KRAS mutant LUAD patients whose tumors harbor high expression of Mucin 5 AC (MUC5AC), a main airway secretory mucin, have significantly lower survival and poor prognosis. Mucins are membrane-bound and secreted hydrophilic proteins essential in clearance of allergens, pathogens, and debris in the lung airway. However, overproduction of mucins, such as MUC5AC, plays roles in the pathogenesis of airway diseases such as chronic obstructive pulmonary disease (COPD) and lung cancer. Given the known effect of mucin polymers on bacterial growth and the immune microenvironment in the context of other airway inflammatory diseases, in this study, our objective is to better understand the MUC5AC-to-microbiome relationship using a lung epithelial specific mouse model of KRAS-mutant LUAD (CC-LR) and test the combinatorial effects of genetic deletion of MUC5AC (CC-LR/Muc5ac−/−) and the pan-microbiome depletion on tumor progression, the lung immune contexture, and microbiome (lung and gut). Thus far, we found that genetic deletion of MUC5AC or pan-microbiome depletion significantly decreases the tumor burden (66% vs 50%, respectively). Additionally, both groups show to have a significantly decreased percentage of tumor-specific Ki67+ cells, a cell proliferation marker, and ERG+ cells, an angiogenesis marker, in comparison to the CC-LR controls. The combination of the lack of MUC5AC with pan-microbiome depletion has also shown to reduce tumor development that needs further investigation. These findings highlight the need to better understand the mechanistic interaction between mucins and the lung/gut microbiome in the context of KRAS-mutant lung cancer which we are currently exploring. It also suggests that their relationship could be a key target for modulation, potentially leading to the development of a minimally invasive alternative preventive and/or treatment modality against KRAS-mutant LUAD. Citation Format: Maria T. Grimaldo, Michael J. Clowers, Avantika Krishna, Katherine E. Larsen, Nastaran Karimi, Yasmina H. Rezai, Arnav Gaitonde, Farbod Khalaj, Haoyue Liu, Florencia McAllister, Seyed Javad Moghaddam. Dissecting the relationship between mucin 5AC and the lung/gut microbiome in the pathogenesis of K-ras mutant lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6675.

Bee Sting Venom as a Viable Therapy for Breast Cancer: A Review Article.

Breast cancer is a kind of aggressive cancer that significantly affects women worldwide, thus making research on alternative and new therapies necessary. The potential impact of bee venom on breast cancer is the main subject of this analysis of this research article. Bee venom has drawn the attention of the world with the help of its constituent ingredients, namely the bioactive compounds, enzymes, and complex blend of proteins. They have a particularly varied chemical makeup and proven anti-cancer capabilities. This is a detailed review demonstrating the components of bee venom and their individual functions in fighting cancer, as well as the results of previously conducted in-vitro and in-vivo research. As described later, bee venom has given positive results in triggering apoptosis, preventing cell migration, inhibiting metastasis and invasion, and suppressing the existing breast cancer cells. It is found to have worked better along with the already existing chemotherapy treatments. These results were also proved with the help of various animal studies that showed reduced tumor development, reduced metastasis, and improved therapeutic effectiveness. Furthermore, certain studies and case reports from all over the world have exhibited consistent results in females affected by breast cancer. This study found that people receiving chemotherapy experienced improved health outcomes and reduced discomfort, with fewer negative side effects. It is important to conduct extensive research on the safety and effectiveness of this treatment because it is yet to be approved. The ideal dosage and administration methods must be explored in clinical trials. Moreover, it is imperative to evaluate the results of any combined treatments with current medications. There should be constant monitoring to prevent any potential side effects. Other important things like allergic reactions and hidden concerns should also be considered.

Thermosensitive Polymeric Nanoparticles for Drug Co-Encapsulation and Breast Cancer Treatment.

Despite advances in breast cancer treatment, there remains a need for local management of noninvasive, low-grade ductal carcinoma in situ (DCIS). These focal lesions are well suited for local intraductal treatment. Intraductal administration supported target site drug retention, improved efficacy, and reduced systemic exposure. Here, we used a poly(N-isopropyl acrylamide, pNIPAM) nanoparticle delivery system loaded with cytotoxic piplartine and an MAPKAP Kinase 2 inhibitor (YARA) for this purpose. For tumor environment targeting, a collagen-binding peptide SILY (RRANAALKAGELYKSILYGSG-hydrazide) was attached to pNIPAM nanoparticles, and the nanoparticle diameter, zeta potential, drug loading, and release were assessed. The system was evaluated for cytotoxicity in a 2D cell culture and 3D spheroids. In vivo efficacy was evaluated using a chemical carcinogenesis model in female Sprague-Dawley rats. Nanoparticle delivery significantly reduced the IC50 of piplartine (4.9 times) compared to the drug in solution. The combination of piplartine and YARA in nanoparticles further reduced the piplartine IC50 (~15 times). Treatment with these nanoparticles decreased the in vivo tumor incidence (5.2 times). Notably, the concentration of piplartine in mammary glands treated with nanoparticles (35.3 ± 22.4 μg/mL) was substantially higher than in plasma (0.7 ± 0.05 μg/mL), demonstrating targeted drug retention. These results indicate that our nanocarrier system effectively reduced tumor development with low systemic exposure.

Synthesis, photophysical properties, anticancer evaluation, and molecular docking studies of new pyrimidine linked 4-arylidene-thiazolidin-4-ones as potent anticancer agents.

The reaction of 4-(chloroacetamido)pyrimidine (1) with ammonium thiocyanate gave 2-(pyrimidin-4-ylimino)thiazolidin-4-one (2), which, when condensed with four substituted benzaldehyde analogues, gave the consequent 5-arylidine-2-(pyrimidin-4-ylimino)thiazolidin-4-ones 3a-d. In addition, the absorbance and fluorescence behaviours of pyrimidinylimino-thiazolidin-4-one hybrids 3a-d in various organic solvents were investigated. The emphasis was on studying UV absorption capacities and the effect of various structural components on photophysical qualities such as the 5-arylidene-2-(pyrimidin-4-ylimino)thiazolidin-4-ones and N,N-dimethylamino tail. The cytotoxic effect of four pyrimidinylimino-thiazolidin-4-one hybrids 3a-d on tumour cell lines (HepG2, HCT-116, PC3, MCF-7) and a normal cell line (WI38) is investigated in this work. The cytotoxicity was measured by comparing the half-maximal inhibitory concentration (IC50 ) to the reference medication, 5-fluorouracil. The findings indicate that these hybrid compounds had varying cytotoxic effects on the cell lines examined; hybrids 3b and 3c demonstrated significant anticancer activity against MCF-7 with IC50 values of 7.53 ±0.43 and 9.17 ±0.31 μM, respectively. The inhibitory efficacy of various synthesized hybrids on the epidermal growth factor receptor (EGFR) kinase was investigated. EGFR is a crucial target in cancer treatment because inhibiting it may reduce tumour development and proliferation. The IC50 value was used to calculate the inhibitory activity, which is the concentration of inhibitor necessary to induce half-maximal inhibition of EGFR kinase activity. In addition, the predicted ADME results show that pyrimidinylimino-thiazolidin-4-one hybrids have good pharmacokinetic properties; hybrid 3d is more lipophilic than the other compounds. It has a medium molecular weight, a small number of hydrogen bond acceptors and donors, and a large number of aromatic heavy atoms. Moreover, molecular docking simulations revealed precise information on the interactions of pyrimidinylimino-thiazolidin-4-one hybrids 3a-d and 5-Fu with their respective protein targets. These interactions point to possible pathways for their biological activities and call for more testing to establish their effectiveness as bioactive molecules or therapeutic candidates.

Influence of the Hypoxia-Activated Prodrug Evofosfamide (TH-302) on Glycolytic Metabolism of Canine Glioma: A Potential Improvement in Cancer Metabolism

The transcription factor hypoxia-inducible factor 1α (HIF-1α) drives metabolic reprogramming in gliomas (GLs) under hypoxic conditions, promoting glycolysis for tumor development. Evofosfamide (EVO) releases a DNA-alkylating agent within hypoxic regions, indicating that it may serve as a hypoxia-targeted therapy. The aim of this study was to investigate the glycolytic metabolism and antitumor effects of EVO in a canine GL model. Our clinical data showed that overall survival was significantly decreased in GL dog patients with higher HIF-1α expression compared to that of those with lower HIF-1α expression, and there was a positive correlation between HIF-1α and pyruvate dehydrogenase kinase 1 (PDK1) expression, suggesting that glycolytic activity under hypoxia conditions may contribute to poor outcomes in canine GL. Our glycolysis assay tests showed that the glycolytic ATP level was higher than the mitochondrial ATP level in three types of canine GL cell lines by activating the HIF-1 signal pathway under hypoxia conditions, resulting in an overall increase in total cellular ATP production. However, treatment with EVO inhibited the glycolytic ATP level in the GL cell lines under hypoxia conditions by targeting HIF-1α-positive cells, leading to decrease in total cellular ATP production. Our in vivo tests showed that EVO significantly reduced tumor development compared to controls and temozolomide in murine GL models. A metabolic analysis demonstrated that EVO effectively suppressed glycolytic metabolism by eliminating HIF-1α-positive cells, suggesting that it may restore metabolism in canine GLs. The evidence presented here supports the favorable preclinical evaluation of EVO as a potential improvement in cancer metabolism.

Stereoselective Synthesis of Sialyl Lewisa Antigen and the Effective Anticancer Activity of Its Bacteriophage Qβ Conjugate as an Anticancer Vaccine

AbstractSialyl Lewisa (sLea), also known as cancer antigen 19‐9 (CA19‐9), is a tumor‐associated carbohydrate antigen. The overexpression of sLea on the surface of a variety of cancer cells makes it an attractive target for anticancer immunotherapy. However, sLea‐based anticancer vaccines have been under‐explored. To develop a new vaccine, efficient stereoselective synthesis of sLea with an amine‐bearing linker was achieved, which was subsequently conjugated with a powerful carrier bacteriophage, Qβ. Mouse immunization with the Qβ‐sLea conjugate generated strong and long‐lasting anti‐sLea IgG antibody responses, which were superior to those induced by the corresponding conjugate of sLea with the benchmark carrier keyhole limpet hemocyanin. Antibodies elicited by Qβ‐sLea were highly selective toward the sLea structure, could bind strongly with sLea‐expressing cancer cells and human pancreatic cancer tissues, and kill tumor cells through complement‐mediated cytotoxicity. Furthermore, vaccination with Qβ‐sLea significantly reduced tumor development in a metastatic cancer model in mice, demonstrating tumor protection for the first time by a sLea‐based vaccine, thus highlighting the significant potential of sLea as a promising cancer antigen.

Stereoselective Synthesis of Sialyl Lewisa Antigen and the Effective Anticancer Activity of Its Bacteriophage Qβ Conjugate as an Anticancer Vaccine.

Sialyl Lewisa (sLea ), also known as cancer antigen 19-9 (CA19-9), is a tumor-associated carbohydrate antigen. The overexpression of sLea on the surface of a variety of cancer cells makes it an attractive target for anticancer immunotherapy. However, sLea -based anticancer vaccines have been under-explored. To develop a new vaccine, efficient stereoselective synthesis of sLea with an amine-bearing linker was achieved, which was subsequently conjugated with a powerful carrier bacteriophage, Qβ. Mouse immunization with the Qβ-sLea conjugate generated strong and long-lasting anti-sLea IgG antibody responses, which were superior to those induced by the corresponding conjugate of sLea with the benchmark carrier keyhole limpet hemocyanin. Antibodies elicited by Qβ-sLea were highly selective toward the sLea structure, could bind strongly with sLea -expressing cancer cells and human pancreatic cancer tissues, and kill tumor cells through complement-mediated cytotoxicity. Furthermore, vaccination with Qβ-sLea significantly reduced tumor development in a metastatic cancer model in mice, demonstrating tumor protection for the first time by a sLea -based vaccine, thus highlighting the significant potential of sLea as a promising cancer antigen.

Targeting transforming growth factor beta (TGF-β) using Pirfenidone, a potential repurposing therapeutic strategy in colorectal cancer

The modulating factors within the tumor microenvironment, for example, transforming growth factor beta (TGF-β), may limit the response to chemo and immunotherapy protocols in colorectal cancer (CRC). In the current study, the therapeutic potential of targeting the TGF-β pathway using Pirfenidone (PFD), a TGF-β inhibitor, either alone or in combination with five fluorouracil (5-FU) has been explored in preclinical models of CRC. The anti-proliferative and migratory effects of PFD were assessed by MTT and wound-healing assays respectively. Xenograft models were used to study the anti-tumor activity, histopathological, and side effects analysis. Targeting of TGF-β resulted in suppression of cell proliferation and migration, associated with modulation of survivin and MMP9/E-cadherin. Moreover, the PFD inhibited TGF-β induced tumor progression, fibrosis, and inflammatory response through perturbation of collagen and E-cadherin. Targeting the TGF-β pathway using PFD may increase the anti-tumor effects of 5-FU and reduce tumor development, providing a new therapeutic approach to CRC treatment.

Ellagic Acid - A Dietary Polyphenol with Anticancer Activity that Deserves More Consideration

Ellagic acid (EA) the principal polyphenol of pomegranate (<em>Punicca granatum</em>) is renowned for its beneficial therapeutic activity in several diseases including cancer. Studies have shown that EA exerts a carcinopreventive effect on many cancer cells, inducing cell cycle arrest and apoptosis, and limiting neovascularization and cell migration. In animal models, EA could reduce tumor development, number and size. Research detected that EA exerts its activity on cancer cells through several signaling pathways, without affecting the function and viability of normal cells. While the investigation of the carcinopreventive effect of EA has been carried out in great detail <em>in vitro</em>, research on animal models and clinical studies is rather scanty, leaving the impression that its potential value as a carcinopreventer is somewhat neglected. This work aims to review the effect of EA on different types of cancer and to attract researchers’ attention to the potential of EA to serve as an adjuvant to anti-cancer drugs.

Polymeric graphene oxide nanoparticles loaded with doxorubicin for combined photothermal and chemotherapy in triple negative breast cancer

Combining photothermal and chemotherapy is an emerging strategy for tumor irradiation in a minimally invasive manner, utilizing photothermal transduction agents and anticancer drugs. The present work developed a 2D carbon nanomaterial graphene oxide (GO)-based nanoplatform that converted to 3D colloidal spherical structures upon functionalization with an amphiphilic polymer mPEG-PLA (1, 0.5/1/2) and entrapped doxorubicin (Dox) physically. The Dox@GO(mPP) (1/0.5) NPs displayed the least particle size (161 nm), the highest stability with no aggregation, the highest Dox loading (6.3 %) and encapsulation efficiency (70 %). The therapeutic efficacy was determined in vitro and in vivo using murine (4 T1) and human triple-negative breast cancer cells (MDA-MB-231), and 4 T1-Luc-tumor bearing mouse models. The results demonstrated that the Dox@GO(mPP) (1/0.5) NPs treatment with laser (+L) (808 nm) was highly efficient in inducing apoptosis, cell cycle arrest (G2/M) phase, significant cytotoxicity, mitochondrial membrane depolarization, ROS generation, and photothermal effect leading to a higher proportion of cell death than free Dox, and Dox@GO(mPP) (1/0.5) NPs (−L). The anticancer studies in mice harboring the 4 T1-Luc tumor showed that combination of Dox@GO(mPP) (1/0.5) NPs (+L) effectively reduced tumor development and decreased lung metastasis. The developed nanoplatform could be a promising combination chemo-photothermal treatment option for triple-negative breast cancer.

Prevalence and influence of overweight and obesity on clinical and epidemiological profile of breast cancer patients in the North of Mato Grosso, Brazil: a retrospective cross-sectional study

Obesity is one of the main preventable risk factors in post-menopausal breast cancer. This retrospective cross-sectional study aimed to demonstrate the clinical and epidemiological profile of breast cancer patients diagnosed in the period from 2013 to 2018 in the North of Mato Grosso, Brazil and to verify the prevalence and influence of overweight and obesity in these patients. Data were collected from patient’s medical records who were diagnosed with breast carcinoma in the Department of Oncology of Santo Antônio’s Hospital, in Sinop-MT. 196 patients were included. 99.5% were women. The majority were married, ≥50 years old (57.7%) and overweight or obese. In the overweight and obesity group the percentage of patients with invasive breast carcinoma were significantly higher when compared with eutrophic group (p=0.03). In all groups the profile of estrogen and progesterone receptors positive and HER-2 negative were more prevalent, however, the frequency of triple negative profile was higher in the overweight (7.1%) and obesity (6.3%) group when compared with control (4.3%) group, as well as the presence of hypertension and diabetes. In conclusion, it was observed a high prevalence of overweight and obesity in breast cancer patients, which contributed to modify the histological type of breast cancer (high prevalence of invasive and lobular carcinomas), increase the frequency of patients in stages 3 and 4, the percentage of triple negative profile and the frequency of other comorbidities, as hypertension and diabetes. Furthermore, metformin, an antidiabetic drug, seems to be contributing to reduce tumor development and improve the clinical profile and prognosis in diabetic breast cancer patients.

SP-8356 (A Verbenone Derivative) Inhibits Proliferation, Suppresses Cell Migration and Invasion and Decreases Tumor Growth of Osteosarcoma: Role of PGC-1α/TFAM and AMPK-Activation.

Osteosarcoma (OS) is an uncommon sarcoma with osteoid formation in conjunction with malignant mesenchymal cells on histological examination. SP-8356 has been reported to exhibit anti-cancer properties in human cancers. However the impact of SP-8356 on OS is largely unknown. The metabolic pathways are coordinated by AMPactivated protein kinase (AMPK), which maintains a balance between the supply and demand of nutrients and energy. This study aimed to investigate effect of SP-8356 on proliferation and apoptosis of OS cells and tumor growth in mice. Furthermore, involvement of PGC-1α/TFAM and AMPK-activation was studied. In the experimental study, Saos-2 and MG63 cells were cultured with SP-8356 for 24 hours and analysed for cellular proliferation using MTT assay. DNA fragmentation was studied using ELISA based kit. Furthermore, transwell chambers assay was used to determine cell migration and cell invasion. Targeted protein expression levels were assessed using western blotting. For in vivo studies, mice (5-6 weeks old) were implanted with either Saos-2 or MG63 cells on dorsal surface subcutaneously and they were administered with SP-8356 (10 mg/kg) for two weeks prior to bone tumor induction. We found that SP-8356 exerted anti-proliferative effects on Saos-2 and MG63 cells. Furthermore, SP-8356 treatment significantly restricted migration and invasion of Saos-2 and MG63 cells. Compared to the control, SP-8356 significantly reduced apoptotic cell death, while it increased PGC-1α and TFAM expressions. Without affecting body weight, SP-8356 significantly reduced tumor development in mice, as compared to the control group. SP-8356 was found to inhibit proliferation, suppressed cells migration and invasion and decreased OS tumor growth. Furthermore, SP-8356 was found to act through PGC-1α/TFAM and AMPK activations. SP-8356 can be therefore used as therapeutic agent for OS treatment.

Cetuximab-conjugated PLGA nanoparticles as a prospective targeting therapeutics for non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is one of the most prevalent cancers diagnosed worldwide, yet managing it is still challenging. The epidermal growth factor receptor (EGFR) exhibits aberrant signalling in a wide range of human cancers, and it is reported to overexpress in most NSCLC cases. The monoclonal antibody [Cetuximab (Cet)] was conjugated onto the surface of the poly (lactide-co-glycolide) (PLGA) nanoparticles which were loaded with docetaxel (DTX) for the development of targeted therapy against lung cancer. This site-specific delivery system exhibited an enhanced cellular uptake in lung cancer cells which overexpress EGFR (A549 and NCI-H23). The nanoparticles also showed better therapeutic effectiveness against NSCLC cells, as evidenced by reduced IC50 values, cell cycle arrest at the G2/M phase, and increased apoptosis. The improved efficacy and in vivo tolerance of Cet-DTX NPs were demonstrated in benzo(a)pyrene (BaP)-induced lung cancer mice model. Histopathological analysis showed that intravenous injection of Cet-DTX NP to mice carrying lung cancer greatly reduced tumour development and proliferation. Comparing Cet-DTX NP to free drug and unconjugated nanoparticles, it also had negligible side effects and improved survival rates. Therefore, Cet-DTX NPs present a promising active targeting carrier for lung tumour-NSCLC-selective treatment.