Modulate Tumor Growth Research Articles

Dietary Risk Factors for Colorectal Cancer and Progress in The Application of Dietary Regulation in Treatment

rectal cancer, a subtype of colorectal cancer, has a significant incidence rate in both men and women, making it a critical area of study. Understanding the impact of daily dietary intake—including protein, fat, vitamins, and trace elements. The fear surrounding cancer stems from its complexity, variability, and the devastating impact it can have on individuals and their families. Cancer’s relentless nature, characterized by the abnormal and unchecked proliferation of cells, underscores the importance of early detection and prevention. The link between unhealthy lifestyle habits and cancer risk underscores the need for individuals to adopt healthier behaviors, such as maintaining a balanced diet, engaging in regular physical activity, and avoiding harmful substances like tobacco and excessive alcohol consumption. This study examines how dietary modifications can reduce the risk of developing Colorectal cancer and their role in supporting treatment, potentially by modulating tumor growth, boosting immune function, and regulating gut flora.

One-Pot multicomponent synthesis of novel pyrazole-linked thiazolyl-pyrazolines: Molecular docking and cytotoxicity assessment on breast and lung cancer cell-lines

Cancer remains a foremost cause of mortality globally, with approximately 20 million new cases and nearly 10 million deaths reported in 2022. Lung and breast cancers are particularly prevalent and deadly, underscoring the critical need for novel therapeutic strategies. This study utilized a one-pot, multicomponent reaction (MCR) approach to synthesize a novel series of pyrazole-linked thiazolyl-pyrazolines. The optimal reaction conditions were determined by systematically varying bases, temperatures, reaction times, and solvents. The best yield was achieved using NaOH as the base at 50 °C with 2 h of stirring in ethanol. The synthesized compounds underwent thorough characterization through IR, mass spectrometry, and NMR spectroscopy. Molecular docking studies indicated strong binding affinities to the ErbB4 kinase, suggesting potential utility as kinase inhibitors. Compounds 5E, 5F, and 5G exhibited significant cytotoxicity against breast cancer cell lines, with IC50 values ranging from 15.79±1.272 to 28.38±1.520 µM. Meanwhile, compounds 5A, 5B, 5D, and 5G demonstrated potent effects against lung cancer cell lines, with IC50 values between 7.945±0.99 and 9.295±1.074 µM. Remarkably, compound 5G showed potent efficacy against both breast and lung cancers, with IC50 values of 28.38±1.520 µM and 9.02±1.07 µM, respectively. This study illuminates the critical role of pyrazole-linked thiazolyl-pyrazoline scaffolds in crafting powerful cancer therapeutics targeting ErbB4 (HER4) kinase. Unique in its dual functionality, ErbB4 can act as either an oncogene or a tumor suppressor in lung and breast cancers, contingent on specific cellular contexts and isoform variations. The dynamic expression of ErbB4 intricately modulates tumor growth and patient outcomes, marking it as a compelling therapeutic target for innovative treatments in both breast and lung cancer.

Spatial interactions modulate tumor growth and immune infiltration

Direct observation of tumor-immune interactions is unlikely in tumors with currently available technology, but computational simulations based on clinical data can provide insight to test hypotheses. It is hypothesized that patterns of collagen evolve as a mechanism of immune escape, but the exact nature of immune-collagen interactions is poorly understood. Spatial data quantifying collagen fiber alignment in squamous cell carcinomas indicates that late-stage disease is associated with highly aligned fibers. Our computational modeling framework discriminates between two hypotheses: immune cell migration that moves (1) parallel or (2) perpendicular to collagen fiber orientation. The modeling recapitulates immune-extracellular matrix interactions where collagen patterns provide immune protection, leading to an emergent inverse relationship between disease stage and immune coverage. Here, computational modeling provides important mechanistic insights by defining a kernel cell-cell interaction function that considers a spectrum of local (cell-scale) to global (tumor-scale) spatial interactions. Short-range interaction kernels provide a mechanism for tumor cell survival under conditions with strong Allee effects, while asymmetric tumor-immune interaction kernels lead to poor immune response. Thus, the length scale of tumor-immune interaction kernels drives tumor growth and infiltration.

IDH2 regulates macrophage polarization and tumorigenesis by modulating mitochondrial metabolism in macrophages

BackgroundTargeting the tumor microenvironment represents an emerging therapeutic strategy for cancer. Macrophages are an essential part of the tumor microenvironment. Macrophage polarization is modulated by mitochondrial metabolism, including oxidative phosphorylation (OXPHOS), the tricarboxylic acid (TCA) cycle, and reactive oxygen species content. Isocitrate dehydrogenase 2 (IDH2), an enzyme involved in the TCA cycle, reportedly promotes cancer progression. However, the mechanisms through which IDH2 influences macrophage polarization and modulates tumor growth remain unknown.MethodsIn this study, IDH2-deficient knockout (KO) mice and primary cultured bone marrow-derived macrophages (BMDMs) were used. Both in vivo subcutaneous tumor experiments and in vitro co-culture experiments were performed, and samples were collected for analysis. Western blotting, RNA quantitative analysis, immunohistochemistry, and flow cytometry were employed to confirm changes in mitochondrial function and the resulting polarization of macrophages exposed to the tumor microenvironment. To analyze the effect on tumor cells, subcutaneous tumor size was measured, and growth and metastasis markers were identified.ResultsIDH2-deficient macrophages co-cultured with cancer cells were found to possess increased mitochondrial dysfunction and fission than wild-type BMDM. Additionally, the levels of M2-associated markers decreased, whereas M1-associated factor levels increased in IDH2-deficient macrophages. IDH2-deficient macrophages were predominantly M1. Tumor sizes in the IDH2-deficient mouse group were significantly smaller than in the wild-type mouse group. IDH2 deficiency in macrophages was associated with inhibited tumor growth and epithelial–mesenchymal transition.ConclusionsOur findings suggest that IDH2 deficiency inhibits M2 macrophage polarization and suppresses tumorigenesis. This study underlines the potential contribution of IDH2 expression in macrophages and tumor microenvironment remodeling, which could be useful in clinical cancer research.

Design, Synthesis and Antitumor Activity of a Novel Class of SHP2 Allosteric Inhibitors with a Furanyl Amide-Based Scaffold.

SHP2 plays a critical role in modulating tumor growth and PD-1-related signaling pathway, thereby serving as an attractive antitumor target. To date, no antitumor drugs targeting SHP2 have been approved, and hence, the search of SHP2 inhibitors with new chemical scaffolds is urgently needed. Herein, we developed a novel SHP2 allosteric inhibitor SDUY038 with a furanyl amide scaffold, demonstrating potent binding affinity (KD = 0.29 μM), enzymatic activity (IC50 = 1.2 μM) and similar binding interactions to SHP099. At the cellular level, SDUY038 exhibited pan-antitumor activity (IC50 = 7-24 μM) by suppressing pERK expression. Furthermore, SDUY038 significantly inhibited tumor growth in both xenograft and organoid models. Additionally, SDUY038 displayed acceptable bioavailability (F = 14%) and half-life time (t1/2 = 3.95 h). Conclusively, this study introduces the furanyl amide scaffold as a novel class of SHP2 allosteric inhibitors, offering promising lead compounds for further development of new antitumor therapies targeting SHP2.

ADAMTS16 Suppression by MicroRNA-25 Leads to Oncogenic Properties in Renal Cell Carcinoma.

Increasing evidence indicates that microRNAs (miRNAs) play a crucial role in modulating tumor growth. This study is centered on investigating the contribution of miR-25 to the progression of Renal Cell Carcinoma (RCC). The investigators examined the expression levels of miR-25 and ADAMTS16 in RCC samples and cell lines. The association between miR-25 and ADAMTS16 was validated via a luciferase reporter assay. Cell viability, apoptosis, migration, and invasion were evaluated utilizing CCK-8 and flow cytometry techniques, while the expression levels of ADAMTS16, β-catenin, GSK-3β, and p-GSK-3β were assessed through western blot analysis. The investigation revealed elevated expression levels of miR-25 in RCC tissues. Subsequently, ADAMTS16 was identified as a target of miR-25. Increased miR-25 levels were associated with decreased expression of ADAMTS16, resulting in enhanced cell viability and diminished apoptosis. Conversely, inhibition of miR-25 led to decreased cell viability, proliferation, and migration. Additionally, the researchers observed that miR-25 triggered the phosphorylation of GSK-3β and β-catenin while leaving the total GSK-3β level unaffected. This study suggests that miR-25 regulates the expression of ADAMTS16 through the Wnt/β-catenin signaling pathway, providing new insights into the cause and potential treatment of RCC.

The potential role of renin angiotensin system in acute leukemia: a narrative review.

Acute leukemias (ALs) are the most common cancers in pediatric population. There are two types of ALs: acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Some studies suggest that the Renin Angiotensin System (RAS) has a role in ALs. RAS signaling modulates, directly and indirectly, cellular activity in different cancers, affecting tumor cells and angiogenesis. Our review aimed to summarize the role of RAS in ALs and to explore future perspectives for the treatment of these hematological malignancies by modulating RAS molecules. The database including Pubmed, Scopus, Cochrane Library, and Scielo were searched to find articles about RAS molecules in ALL and in pediatric patients. The search terms were "RAS", "Acute Leukemia", "ALL", "Angiotensin-(1-7)", "Pediatric", "Cancer", "Angiotensin II", "AML". In the bone marrow, RAS has been found to play a key role in blood cell formation, affecting several processes including apoptosis, cell proliferation, mobilization, intracellular signaling, angiogenesis, fibrosis, and inflammation. Local tissue RAS modulates tumor growth and metastasis through autocrine and paracrine actions. RAS mainly acts via two molecules, Angiotensin II (Ang II) and Angiotensin (1-7) [Ang-(1-7)]. While Ang II promotes tumor cell growth and stimulates angiogenesis, Ang-(1-7) inhibits the proliferation of neoplastic cells and the angiogenesis, suggesting a potential therapeutic role of this molecule in ALL. The interaction between ALs and RAS reveals a complex network of molecules that can affect the hematopoiesis and the development of hematological cancers. Understanding these interactions could pave the way for innovative therapeutic approaches targeting RAS components.

In Vivo Combined Photoacoustic Imaging and Photothermal Treatment of HPV‐Negative Head and Neck Carcinoma with NIR‐Responsive Non‐Persistent Plasmon Nano‐Architectures

AbstractThe combination of photoacoustic imaging (PAI) and photothermal therapy (PTT) is an attractive approach in cancer management due to the non‐invasive features combined with real‐time imaging and selective tissue damage by non‐ionizing radiation. This approach is especially appealing for Head and Neck Squamous Cell Carcinoma (HNSCC) management, where up to 40% of patients require modifications of the treatment regimen. On the other hand, most of the agents developed for PAI/PTT suffer from persistence or re‐shaping issues. Here, a unique non‐persistent plasmon nano‐architecture (tNAs‐IRDye) is presented that simultaneously acts as a contrast agent for PAI and as a photothermal transducer for PTT. The tNAs‐IRDye are fully characterized and evaluated in vitro and ex vivo, and their performance as theranostic agents is assessed in HPV‐negative HNSCC murine models. A significant modulation of tumor growth is obtained in vivo upon intratumoral injection of tNAs‐IRDye and subsequent NIR irradiation compared to the solely irradiated control. The outcomes of this study exhibit a noteworthy potential to foster the development of innovative clinical strategies for the management of HPV‐negative head and neck carcinoma.

Spatial interactions modulate tumor growth and immune infiltration.

Lenia, a cellular automata framework used in artificial life, provides a natural setting to implement mathematical models of cancer incorporating features such as morphogenesis, homeostasis, motility, reproduction, growth, stimuli response, evolvability, and adaptation. Historically, agent-based models of cancer progression have been constructed with rules that govern birth, death and migration, with attempts to map local rules to emergent global growth dynamics. In contrast, Lenia provides a flexible framework for considering a spectrum of local (cell-scale) to global (tumor-scale) dynamics by defining an interaction kernel governing density-dependent growth dynamics. Lenia can recapitulate a range of cancer model classifications including local or global, deterministic or stochastic, non-spatial or spatial, single or multi-population, and off or on-lattice. Lenia is subsequently used to develop data-informed models of 1) single-population growth dynamics, 2) multi-population cell-cell competition models, and 3) cell migration or chemotaxis. Mathematical modeling provides important mechanistic insights. First, short-range interaction kernels provide a mechanism for tumor cell survival under conditions with strong Allee effects. Next, we find that asymmetric interaction tumor-immune kernels lead to poor immune response. Finally, modeling recapitulates immune-ECM interactions where patterns of collagen formation provide immune protection, indicated by an emergent inverse relationship between disease stage and immune coverage.

Abstract PO4-26-03: Collagen deposition in the breast cancer microenvironment correlates with ER-positive, Histological Grade I and low proliferation tumors

Abstract Breast cancer is a highly heterogeneous neoplasm both at the molecular and histological levels. Breast cancer cells present extremely high levels of genetic heterogeneity concerning mutational burden, copy number variations (CNVs) and epigenetic signatures. While this level of heterogeneity is well established, other layers of heterogeneity, such as tumor microenvironment (TME) heterogeneity, remains to be elucidated. TME in breast cancer is an intricate network encompassing tumor cells, “normal” stroma cells such as fibroblasts, lymphocytes, and a plethora of extracellular matrix (ECM) proteins produced either by cancer cells or stromal cells. These three interacting modules (cancer cells, stromal cells and the ECM) can affect each other in a reciprocal way with profound effects on patient prognosis and treatment outcome. Major protein components of the TME are ECM-related collagens, that are produced by breast cancer cells or/and cancer associated fibroblasts. The ECM collagen in breast cancer, that is mainly composed of collagen types I, III and V, has multiple roles modulating tumor growth, cancer cell invasiveness and metastasis. Here by Masson Trichrome and Immunohistochemistry/Aniline Blue co-stains for collagen fibers and proliferation index (Ki-67) in breast cancer FFPE tissue samples, we identified a statistically significant correlation between collagen deposition and Estrogen Receptor (ER)) status (p< 0.05, proliferation index (p< 0.01) and histological Grade (p=0.01). Further, in order to expand and support our findings, we analyzed the METABRIC study through the cBioPortal hub in relevance to collagen genes and multiple clinicopathological parameters. We were able to identify a coregulated subset of collagen genes that presented a positive correlation with ER-positive (p< 10−10, q< 10−10), low proliferating (p< 10−10, q< 10−10), Grade I (p=2.35e−9, q< 1.07e−8) group of breast cancer cases, supporting our findings. In addition, analyses of collagen-high patients from the METABRIC study indicated that most cases presented with PIK3CA, CBFB and MAP3K1 mutations, while through the g.Profiler software we identified collagen-high relevant gene signatures that encompassed gene ontology terms related to tissue development, extracellular matrix organization and cell migration. Collectively, our preliminary results indicate that ECM collagen deposition in breast cancer is characteristic of low-grade tumors with a better differentiation and possibly with a favorable prognosis. Citation Format: Angelos Koutras, Chaido Sirinian, Olga Bogri, Martha Nifora, Maria Theakou, Soren Degn, Ioannis Mouratidis, Dimitrios Chaniotis, Stavros Peroukidis, Ilias Georgakopoulos-Soares, Anastasios Papanastasiou. Collagen deposition in the breast cancer microenvironment correlates with ER-positive, Histological Grade I and low proliferation tumors [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-26-03.

An inflammation-related subtype classification for analyzing tumor microenvironment and clinical prognosis in colorectal cancer.

The inflammatory response plays an essential role in the tumor microenvironment (TME) of colorectal cancer (CRC) by modulating tumor growth, progression, and response to therapy through the recruitment of immune cells, production of cytokines, and activation of signaling pathways. However, the molecular subtypes and risk score prognostic model based on inflammatory response remain to be further explored. Inflammation-related genes were collected from the molecular signature database and molecular subtypes were identified using nonnegative matrix factorization based on the TCGA cohort. We compared the clinicopathological features, immune infiltration, somatic mutation profile, survival prognosis, and drug sensitivity between the subtypes. The risk score model was developed using LASSO and multivariate Cox regression in the TCGA cohort. The above results were independently validated in the GEO cohort. Moreover, we explored the biological functions of the hub gene, receptor interacting protein kinase 2 (RIPK2), leveraging proteomics data, in vivo, and in vitro experiments. We identified two inflammation-related subtypes (inflammation-low and inflammation-high) and have excellent internal consistency and stability. Inflammation-high subtype showed higher immune cell infiltration and increased sensitivity to common chemotherapeutic drugs, while inflammation-low subtype may be more suitable for immunotherapy. Besides, the two subtypes differ significantly in pathway enrichment and biological functions. In addition, the 11-gene signature prognostic model constructed from inflammation-related genes showed strong prognostic assessment power and could serve as a novel prognostic marker to predict the survival of CRC patients. Finally, RIPK2 plays a crucial role in promoting malignant proliferation of CRC cell validated by experiment. This study provides new insights into the heterogeneity of CRC and provides novel opportunities for treatment development and clinical decision making.

Extracellular Vesicle-Related Non-Coding RNAs in Hepatocellular Carcinoma: An Overview.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer. It is a major public health problem worldwide, and it is often diagnosed at advanced stages, when no effective treatment options are available. Extracellular vesicles (EVs) are nanosized double-layer lipid vesicles containing various biomolecule cargoes, such as lipids, proteins, and nucleic acids. EVs are released from nearly all types of cells and have been shown to play an important role in cell-to-cell communication. In recent years, many studies have investigated the role of EVs in cancer, including HCC. Emerging studies have shown that EVs play primary roles in the development and progression of cancer, modulating tumor growth and metastasis formation. Moreover, it has been observed that non-coding RNAs (ncRNAs) carried by tumor cell-derived EVs promote tumorigenesis, regulating the tumor microenvironment (TME) and playing critical roles in the progression, angiogenesis, metastasis, immune escape, and drug resistance of HCC. EV-related ncRNAs can provide information regarding disease status, thus encompassing a role as biomarkers. In this review, we discuss the main roles of ncRNAs present in HCC-derived EVs, including micro(mi) RNAs, long non-coding (lnc) RNAs, and circular (circ) RNAs, and their potential clinical value as biomarkers and therapeutic targets.

ZMIZ1 enhances ERα-dependent expression of E2F2 in breast cancer.

The estrogen receptor-α (ER) drives 75% of breast cancers. On activation, the ER recruits and assembles a 1-2 MDa transcriptionally active complex. These complexes can modulate tumour growth, and understanding the roles of individual proteins within these complexes can help identify new therapeutic targets. Here, we present the discovery of ER and ZMIZ1 within the same multi-protein assembly by quantitative proteomics, and validated by proximity ligation assay. We characterise ZMIZ1 function by demonstrating a significant decrease in the proliferation of ER-positive cancer cell lines. To establish a role for the ER-ZMIZ1 interaction, we measured the transcriptional changes in the estrogen response post-ZMIZ1 knockdown using an RNA-seq time-course over 24 h. Gene set enrichment analysis of the ZMIZ1-knockdown data identified a specific delay in the response of estradiol-induced cell cycle genes. Integration of ENCODE data with our RNA-seq results identified that ER and ZMIZ1 both bind the promoter of E2F2. We therefore propose that ER and ZMIZ1 interact to enable the efficient estrogenic response at subset of cell cycle genes via a novel ZMIZ1-ER-E2F2 signalling axis. Finally, we show that high ZMIZ1 expression is predictive of worse patient outcome, ER and ZMIZ1 are co-expressed in breast cancer patients in TCGA and METABRIC, and the proteins are co-localised within the nuclei of tumour cell in patient biopsies. In conclusion, we establish that ZMIZ1 is a regulator of the estrogenic cell cycle response and provide evidence of the biological importance of the ER-ZMIZ1 interaction in ER-positive patient tumours, supporting potential clinical relevance.

Collagen Lattice Model, Populated with Heterogeneous Cancer-Associated Fibroblasts, Facilitates Advanced Reconstruction of Pancreatic Cancer Microenvironment.

Pancreatic ductal adenocarcinoma (PDAC) is a solid-tumor malignancy. To enhance the treatment landscape of PDAC, a 3D model optimized for rigorous drug screening is essential. Within the PDAC tumor microenvironment, a dense stroma comprising a large extracellular matrix and cancer-associated fibroblasts (CAFs) is well-known for its vital role in modulating tumor growth, cellular heterogeneity, bidirectional paracrine signaling, and chemoresistance. In this study, we employed a fibroblast-populated collagen lattice (FPCL) modeling approach that has the ability to replicate fibroblast contractility in the collagenous matrix to build dense stroma. This FPCL model allows CAF differentiation by facilitating multifaceted cell-cell interactions between cancer cells and CAFs, with the differentiation further influenced by mechanical forces and hypoxia carried within the 3D structure. Our FPCL models displayed hallmark features, including ductal gland structures and differentiated CAFs with spindle shapes. Through morphological explorations alongside in-depth transcriptomic and metabolomic profiling, we identified substantial molecular shifts from the nascent to mature model stages and potential metabolic biomarkers, such as proline. The initial pharmacological assays highlighted the effectiveness of our FPCL model in screening for improved therapeutic strategies. In conclusion, our PDAC modeling platform mirrors complex tumor microenvironmental dynamics and offers an unparalleled perspective for therapeutic exploration.

Abstract 3085: Cholesterol sulfotransferase SULT2B1b suppresses prostate tumor growth and regulates cholesterol metabolism in prostate cancer cells

Abstract Prostate cancer (PrCa) ranks as the second leading cause of cancer death in males in the United States. The 5-year survival rate drops drastically from 100% in patients with early-stage confined PrCa to 30% in those with advanced metastatic PrCa. Patients with advanced PrCa face limited long-term treatment options, often developing resistance to androgen-deprivation therapy and chemotherapy. Identifying and targeting pivotal molecules or pathways in advanced PrCa can substantially enhance patient survival. Our previous findings unveiled an inverse relationship between cholesterol sulfotransferase SULT2B1b and the severity of human PrCa. The highest levels of SULT2B1b were observed in the epithelium of normal prostates, with a progressive decrease as cancer advanced. The lowest levels were found in the most advanced metastatic samples. This study aims to determine whether SULT2B1b performs any active and critical functions in PrCa progression and assess whether it regulates cholesterol metabolism in PrCa cells. We generated SULT2B1b knockout (KO) clones from mouse PrCa cell line MyC-CaP and established inducible SULT2B1b overexpression lines from androgen receptor (AR)+ LNCaP and AR- PC3 human PrCa cells. These modified mouse and human cells were implanted into syngeneic FVB mice and immunodeficient NRG mice, respectively. MyC-CaP KO cells formed significantly larger tumors compared to parental cells, accompanied by reduced survival rates. In contrast, both LNCaP and PC3 tumors with induced SULT2B1b exhibited smaller sizes compared to control tumors, leading to improved survival rates. To investigate the impact of SULT2B1b on cholesterol metabolism and the underlying mechanisms behind SULT2B1b-regulated tumor growth, we examined genes related to lipoprotein signaling and cholesterol metabolism in MyC-CaP SULT2B1b KO cells using PCR arrays and qRT-PCR. Genes responsible for cholesterol uptake and synthesis (e.g. Ldlr, Cyp51, Sorl1, Pcsk9 and Insig1) were among the top upregulated genes, while genes controlling cholesterol efflux (e.g. Abca1) and esterification (e.g. Soat1 and Soat2) were downregulated in KO cells. Consistent with these gene expression patterns, intracellular cholesterol levels significantly increased in KO cells. Interestingly, although SULT2B1b did not affect cell proliferation under normal conditions, KO cells exhibited faster growth than parental cells when cultured in lipid-depleted or charcoal-stripped media, possibly due to altered intracellular cholesterol levels. In summary, these findings align with the expression pattern of SULT2B1b in human PrCa, demonstrating that SULT2B1b inhibits tumor growth, whereas its removal promotes tumor growth. This suggests SULT2B1b may serve as a critical tumor suppressor in PrCa. Its modulation of tumor growth may occur through the regulation of cholesterol metabolism. Citation Format: Jiang Yang, Dilinaer Wusiman, Timothy L. Ratliff. Cholesterol sulfotransferase SULT2B1b suppresses prostate tumor growth and regulates cholesterol metabolism in prostate cancer cells [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 3085.

Abstract 7235: Exosome-mediated delivery of siRNA against NRF2 for treatment of lung cancer

Abstract Nuclear factor erythroid 2-related factor 2 (NRF2) activation is the third most common genetic event in lung adenocarcinomas (25%), primarily induced by loss of function mutations in its master regulator, KEAP1. NRF2 is a key modulator of tumor growth and metabolic reprogramming, resulting in resistance to traditional chemotherapeutics. Due to its role in modulating oxidative stress, sustained NRF2 overexpression promotes disease progression, particularly in lung cancer. This study demonstrates the effective knockdown of NRF2 expression by small interfering RNA (siRNA) delivered via a nanoplatform based on bovine colostrum exosomes to lung cancer cells, both in vitro and in vivo, resulting in inhibition of proliferation and disease progression. Bovine colostrum is an abundant source of exosomes. Here we present a novel exosome-polyethyleneimine matrix (EPM) to deliver therapeutic siRNA targeting NRF2 expression (siNRF2). Specific delivery of the EPM to tumors was further enhanced via folic acid (FA) functionalization, targeting folate receptors highly expressed on lung tumor cells. Exosomes were isolated from standardized colostrum powder through rehydration and differential centrifugation, and characterized by size, polydispersity index (pdi) and surface charge by Zetasizer, and the presence of surface protein markers by Western blot. siNRF2 was loaded onto the EPM, and the resultant EPM-siNRF2 complex was harvested by PEG precipitation. Entrapment efficiency was determined using 5’-32P-labeled siRNA as a tracer. The gene knockdown efficiency of EPM-siNRF2 was assessed in lung cancer (A549) cells by Western blot and the therapeutic effect of target knockdown in vitro was confirmed by analyzing the corresponding modulation in downstream targets of NRF2. The anti-tumor effect was then evaluated using both subcutaneous and orthotropic lung tumor models in immunocompromised mice. The EPM exhibited high entrapment efficiency (>90%) of siNRF2 (up to 20 μg) and protected entrapped siRNA from enzymatic degradation upon exposure to RNases. EPM-siNRF2 dose dependently decreased NRF2 expression in A549 lung cancer cells. When formulated with FA-functionalized exosomes, EPM-siNRF2 inhibited subcutaneous and orthotropic lung tumors in mice by over 57% (p<0.01) and 84% (p<0.001), respectively. Mice treated with FA-EPM-siNRF2 showed decreased levels of NRF2 in tumor tissue, but not in the adjacent lung, indicating FA-mediated targeted delivery. This bovine colostrum exosome based EPM technology showcases high efficiency in gene knockdown and lung tumor inhibition, providing a promising nanoplatform for delivering nucleic acid therapeutics to the tumor site. Funding: Supported from the NIH grant R44 CA-221487, 3P Biotechnology Contract and, in part, Agnes Brown Duggan Endowment. Keywords: Bovine colostrum, Exosomes, NRF2, siRNA, Lung cancer. Citation Format: Raghuram Kandimalla, Margaret Wallen, Jeyaprakash Jeyabalan, Disha N. Moholkar, Wendy Spencer, Ramesh C. Gupta, Farrukh Aqil. Exosome-mediated delivery of siRNA against NRF2 for treatment of 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 7235.

Abstract 5982: Cannabinoid derived product is a potential novel therapeutic for papillary thyroid carcinoma

Abstract Papillary thyroid carcinoma (PTC) is the most common thyroid cancer that usually affects women ages 20-50, presenting as an asymptomatic neck mass. Total or partial thyroidectomy shows an excellent prognosis; however, investigation into non-invasive therapeutics with minimal adverse effects is ongoing. In addition to driver mutations such as BRAF, RAS, and RET-PTC, p53 mutations have also been identified. The loss of function of p53 is linked to cancer progression from PTC to poorly differentiated thyroid cancer and possibly anaplastic thyroid cancer. The disruption of the Wnt cell pathway is documented to be related to malignant cell transformation in thyroid cancer, leading to remarkable upregulation of c-myc, resulting in enhancement of tumor growth and drug resistance. Previous studies revealed that upregulation of BCL-2 occurs at later stages of tumorigenesis and is involved in chemotherapy resistance in some forms of cancer. In PTC, the downregulation of BCL-2 is linked to its favorable prognosis. Cannabinoid Derived Products (CDPs) have shown effectiveness in inhibiting different types of cancer, with research dating back to the 1970s. Ongoing in-vitro and in-vivo studies show that CDPs can effectively modulate tumor growth. However, the apoptotic properties of CDPs are largely dependent on the cancer type and drug dose/concentration. Our investigation focuses on CDPs and its potential anti-cancer effects by modulation of gene expression. We investigated its effects on gene expression of p53, c-Myc, and BCL-2 in K1 cell line which consists of PTC cells obtained from metastasis of well-differentiated PTC. K1 PTC cells (1 x106 cells/mL) were cultured with CDPs and incubated at 37°C under 5% CO2 for 24 and 48 hours. Cells were harvested, and cell viability was determined via trypan blue exclusion assay. Using qRT-PCR, we determined the effects of CDPs on the expression of TP53, c-Myc, and BCL-2. Results show that the CDPs decreased the viability (p< 0.001) of K1 PTC cells in a dose and time-dependent manner. Within 24 hours, the cannabinoid-containing product increased the gene expression of TP53 (p < 0.01) and decreased the gene expression of BCL-2 (p< 0.01) and c-Myc (p< 0.05) in K1 PTC cells. The results suggest that CDPs interact as a potential regulator in cancer with the upregulation of p53 and downregulation of BCL-2 and c-Myc. Further in-vitro and in-vivo studies are needed to understand the mechanism and therapeutic potential of cannabinoid-containing products in papillary thyroid cancer. Citation Format: Carolina Taico, Ibrahim Musa, Fariba Ardalani, Anish Maskey, Nan Yang, Joseph Breslin, Raj Tiwari, Jan Geliebter, Xiu Min Li. Cannabinoid derived product is a potential novel therapeutic for papillary thyroid carcinoma [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 5982.

Abstract 5932: Immunomodulatory and anticancer effects of cyclical gemcitabine/nab-paclitaxel with continuous dosing of tinodasertib (AUM001) in preclinical KPC model

Abstract Background: PDAC has a poor 5-year survival outcome, and its aggressiveness limits the treatment efficacy. This is often driven by dysregulated mRNA translation of pro-tumorigenic genes that contribute to the proliferative and metastatic nature of PDAC. Synergizing chemotherapy with inhibition of oncogenic mRNA translation via MNK-coupled phosphorylation of eIF4E could potentially be viable. To support this hypothesis, we combined AUM001, a MNK inhibitor, with gemcitabine (GEM)-nab-paclitaxel (nPTX) with the aim of modulating tumor growth kinetics and maintaining it in a stable state. Methods: PDAC KPC cells were subcutaneously inoculated into BL6 mice and randomized to five treatment groups. Chemotherapy involved administering GEM-nPTX intraperitoneally on days 1, 4, and 8. AUM001 was given orally daily for 8 days for both mono- and combination therapies and was further administered for an additional 3 weeks in continuous dosing group. IHC staining for Ki67, MVD, and stromal markers was conducted on FFPE sections as well as Western blot analysis of tumor tissues. Apoptosis was assessed using Annexin V. Immune profiling were evaluated by flow cytometric assay and GeoMx spatial transcriptomics analysis. Results: The administration of GEM-nPTX and AUM001 (GPA) or GEM-nPTX alone (GP) led to an average 38% reduction in net tumor growth with no observed toxicity. Notably, continuous dosing of AUM001 (GPAc) following the cessation of chemotherapy exhibited a 45% decrease in net tumor growth (P=0.056) when compared to GPA. EIF4E phosphorylation and Myc expression were markedly inhibited in GPAc with lower recovery rate for MVD as compared to the GPA group at EOO (P < 0.05). A higher fraction of apoptotic cells was detected in the GPAc group at EOO than GPA group in reference to GP group (P=NS). GPA also resulted in a lower frequency of NK cells which was partially lifted in GPAc during observation phase. Macrophages which are known to have a pro-tumor role initially increased in frequency upon addition of AUM during chemotherapy, but subsequently decreased during observation. These modulations in infiltrating immune cells correlated with the differential expression of several immune related genes in the tumor associated CD45+ immune cells, revealed by the GeoMx analysis. Conclusions: This study showed that sustained administration of AUM001 alongside cyclical GEM-nPTX is well-tolerated with good anti-tumor outcomes. Additionally, the maintenance of AUM001 may enhance the decrease in pro-oncogenic protein expression and remodel the tumor immune microenvironment. In conclusion, continuous dosing of AUM001 following the cessation of chemotherapy could stabilize the tumor state, potentiating its application in clinical settings as maintenance therapy for PDAC. Citation Format: Wai Fook Leong, Sze Sing Lee, Jyue Yuan Lim, Siwen Sylvialin Chen, Fara Faye Desacola Regis, Romana Vidergar, Subhra K. Biswas, Balram Chowbay. Immunomodulatory and anticancer effects of cyclical gemcitabine/nab-paclitaxel with continuous dosing of tinodasertib (AUM001) in preclinical KPC 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 5932.

GAS41 modulates ferroptosis by anchoring NRF2 on chromatin

YEATS domain-containing protein GAS41 is a histone reader and oncogene. Here, through genome-wide CRISPR-Cas9 screenings, we identify GAS41 as a repressor of ferroptosis. GAS41 interacts with NRF2 and is critical for NRF2 to activate its targets such as SLC7A11 for modulating ferroptosis. By recognizing the H3K27-acetylation (H3K27-ac) marker, GAS41 is recruited to the SLC7A11 promoter, independent of NRF2 binding. By bridging the interaction between NRF2 and the H3K27-ac marker, GAS41 acts as an anchor for NRF2 on chromatin in a promoter-specific manner for transcriptional activation. Moreover, the GAS41-mediated effect on ferroptosis contributes to its oncogenic role in vivo. These data demonstrate that GAS41 is a target for modulating tumor growth through ferroptosis. Our study reveals a mechanism for GAS41-mediated regulation in transcription by anchoring NRF2 on chromatin, and provides a model in which the DNA binding activity on chromatin by transcriptional factors (NRF2) can be directly regulated by histone markers (H3K27-ac).

Assessment of iridoids and their similar structures as antineoplastic drugs by in silico approach

Iridoids commonly found in plants as secondary metabolites have been reported to possess significant biological activities such as anticancer, antioxidant, hypoglycemic, antimicrobial etc. The strong interactions of iridoids with cyclic-dependent kinase 8 (CDK8) protein could show inhibitory effects that could modulate tumour growth. From the molecular docking calculations, some iridoids interacted effectively with the target CDK8 protein (PDB ID: 5ICP) with better binding affinities of −9.1, −9.0, −9.0, −8.9 kcal/mol, than that observed for the native ligand with −8.7 kcal/mol and for the reference compound gemcitabine with −6.9 kcal/mol. The GI50 values (<5 μM) obtained from graph-based signatures showed activity in breast, colon, leukaemia, and renal cancer cell lines. The IC50 predictions as CDK2 inhibitors were greater than 10 µM with type I non-allosteric binding mode. The stability analysis of protein-ligand complex from 125 ns long molecular dynamics simulations showed moderately smooth trajectories and RSMD value around 5 Å for the docked ligands. The binding free energy changes up to −47.65 ± 5.97 kcal/mol from MMGBSA method and −30.33 ± 5.40 kcal/mol from MMPBSA method hinted at the spontaneous nature of the complex formation. Furthermore, geometrical evaluators like RMSF, Rg, SASA, and hydrogen bond count also corroborated with the structural stability of the complexes and the capacity of hit molecules to inhibit the target, indicating its therapeutic potential against cancer. The toxicity and drug-likeness from ADMET predictions suggested experimental verification and that the proposed candidates could be employed for further trials in the development of safer and more effective anticancer drugs. Communicated by Ramaswamy H. Sarma