Drug Target For Treatment Of Cancer Research Articles

Deubiquitination of CDC6 by OTUD6A promotes tumour progression and chemoresistance

BackgroundCDC6 is an oncogenic protein whose expression level fluctuates during the cell cycle. Although several E3 ubiquitin ligases responsible for the ubiquitin-mediated proteolysis of CDC6 have been identified, the deubiquitination pathway for CDC6 has not been investigated.MethodsThe proteome-wide deubiquitinase (DUB) screening was used to identify the potential regulator of CDC6. Immunofluorescence, protein half-life and deubiquitination assays were performed to determine the protein stability of CDC6. Gain- and loss-of-function experiments were implemented to analyse the impacts of OUTD6A-CDC6 axis on tumour growth and chemosensitivity in vitro. N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced conditional Otud6a knockout (CKO) mouse model and tumour xenograft model were performed to analyse the role of OTUD6A-CDC6 axis in vivo. Tissue specimens were used to determine the association between OTUD6A and CDC6.ResultsOTUD6A interacts with, depolyubiquitinates and stabilizes CDC6 by removing K6-, K33-, and K48-linked polyubiquitination. Moreover, OTUD6A promotes cell proliferation and decreases sensitivity to chemotherapy by upregulating CDC6. CKO mice are less prone to BCa tumorigenesis induced by BBN, and knockdown of OTUD6A inhibits tumour progression in vivo. Furthermore, OTUD6A protein level has a positive correlation with CDC6 protein level, and high protein levels of OTUD6A and CDC6 are associated with poor prognosis in patients with bladder cancer.ConclusionsWe reveal an important yet missing piece of novel DUB governing CDC6 stability. In addition, our findings propose a model for the OTUD6A-CDC6 axis that provides novel insights into cell cycle and chemosensitivity regulation, which may become a potential biomarker and promising drug target for cancer treatment.

Ribosomal protein S6 kinase 2 (RPS6KB2) is a potential immunotherapeutic target for cancer that upregulates proinflammatory cytokines.

Cancer is still a leading cause of mortality. Over the years, cancer therapy has undergone significant advances driven by advancements in science and technology. A promising area of drug discovery in this field involves the development of therapeutic targets for cancer treatment. The urgent need to identify new pharmacological targets arises from the impact of tumor resistance on the effectiveness of current medications. Specifically, the RPS6KB2 gene on chromosome 11 has been implicated in cell cycle regulation and exhibits higher expression levels in tumor tissue. Given this association, there is a potential for this gene to serve as a target for cancer treatment. We conducted an analysis using the GTEx, TCGA, and CCLE databases to explore the relationship between RPS6KB2 and immune infiltration, the tumor microenvironment (TME), microsatellite instability (MSI), and more. Cell proliferation was assessed using EDU detection, while cell invasion and migration were evaluated via wound healing and Transwell assays. Additionally, western blot analysis was employed to measure expression of Bax, Bcl-2, MMP2, MMP9, PCNA, and proinflammatory factors. Through data analysis and molecular biology methods, our study carefully examined the potential role of RPS6KB2 in cancer therapy. The data revealed that RPS6KB2 is aberrantly expressed in most cancers and is associated with poor prognosis. Further analysis indicated its involvement in cancer cell apoptosis and migration, as well as its role in cancer immune processes. We validated the significance of RPS6KB2 in hepatocellular carcinoma (HCC), highlighting its capacity to upregulate proinflammatory cytokines. Our research indicates that RPS6KB2 is a prognostic biomarker associated with immune infiltration in cancer that can affect antitumor immunity by increasing secretion of proinflammatory factors, providing a potential drug target for cancer treatment.

Synthesis and biological evaluation of novel bi-gold mitocans in lung cancer cells.

Mitochondria are promising drug target for cancer treatment. We previously demonstrated that a bi-gold compound BGC2a was more potent than the mono-gold drug auranofin in suppressing cancer cells due to increased gold atom number that led to higher drug accumulation in and thereby inhibition of mitochondria. To exploit the potential of this new strategy, we further designed and synthesized a series of bi-gold mitocans, the compounds targeting mitochondria. The results showed that most of the newly synthesized mitocans exhibited obviously lower IC50 than auranofin, an old drug that is repurposed in clinical trials for cancer treatment. The best mitocan C3P4 was nearly 2-fold more potent than BGC2a in human non-small cell lung cancer A549 cells and mantle cell lymphoma Jeko-1 cells, exhibiting substantial colony formation-suppressing and tumor-suppressing effects in A549 cells xenograft model. C3P4 induced apoptosis in a dose-dependent manner and arrested cell cycle at G0/G1 phase. The mechanistic study showed that C3P4 significantly increased the global reactive oxygen species and mitochondrial superoxide level, and reduced the mitochondrial membrane potential. C3P4 preferentially accumulated in mitochondria as measured by the gold content and substantially inhibited oxygen consumption rate and ATP production. These results further validated our hypothesis that targeting mitochondria would be promising to develop more potent anticancer agents. C3P4 may be further evaluated as a drug candidate for lung cancer treatment.

Abstract LB_B03: Lentivirus-induced ALDH1L1 knock-in inhibits proliferation and migration and activates apoptosis of human A549 lung adenocarcinoma cells

Abstract Lung Adenocarcinoma (LUAD), one of the most common life-threatening malignancies worldwide, represents about 40% of all lung cancers. Human Protein Atlas and OncoLnc database indicate that LUAD tumors have significantly lower mRNA levels of ALDH1L1 (10-formyltetrahydrofolate dehydrogenase, a regulatory enzyme in folate metabolism) than in non-tumor lung tissues. Moreover, higher intratumoral ALDH1L1 expression in LUAD patients correlates with better clinical outcome. These findings raise the question of whether ALDH1L1 knock-in can have a therapeutic effect. As the first step in addressing this question, we knocked-in ALDH1L1 in A549 lung adenocarcinoma cells using the lentiviral delivery. Of note, as most cancer cell lines, A549 cells do not express ALDH1L1. We have constructed the lentiviral vector carrying ALDH1L1 and green fluorescent protein (EGFP) cDNA under the control of CMV and SV40 promoters, respectively. Recombinant lentivirus (Lv-ALDH1L1), generated and packed in HEK293 cells, was used to transduce A549 cells. The efficiency of the transduction was evaluated by monitoring the GFP expression using the fluorescence microscopy of live cells. The expression of ALDH1L1 was determined at the mRNA and protein levels by RT-PCR and western blot assays, accordingly. Our results showed a significant expression of ALDH1L1 mRNA and protein expression in A549 cells carrying the Lv-ALDH1L1 vector. The lentivirus-induced ALDH1L1 elevation produced strong inhibitory effects on cellular proliferation and colony formation under nutrient-rich conditions. Of note, it also induced alterations of cell morphology. To explore the underlying mechanisms of these effects, apoptosis-related markers were examined. Elevated levels of the signal transduction pathways involved in initiating apoptosis; BID, BAX, and BAK in A549 cells transduced with Lv-ALDH1L1 were observed. Furthermore, using the wound healing assay, we showed that ALDH1L1 strongly suppresses the migratory ability of A549 cells. Interestingly, this effect was associated with a robust down-regulation of the actin depolymerizing factor cofilin and its phosphorylated form. Overall, this study demonstrated that lentiviral ALDH1L1 knock-in produces strong anti-tumorigenesis effects in A549 cells. This further suggests that ALDH1L1 should be investigated as a therapeutic drug target in cancer treatment. Citation Format: Amira Abdellatef, Sergey Krupenko. Lentivirus-induced ALDH1L1 knock-in inhibits proliferation and migration and activates apoptosis of human A549 lung adenocarcinoma cells [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr LB_B03.

Investigation of the inhibitory behavior of XFE and mitoxantrone molecules in interaction with AKT1 protein: a molecular dynamics simulation study.

The PI3K/Akt/mTOR pathway is one of the important pathways in many cancers. Akt is a serine-threonine kinase protein identified as a drug target for cancer treatment. Therefore, anticancer drugs are essential therapeutic targets for this pathway. In the current study, the inhibitory effect of two anticancer molecules, XFE and mitoxantrone, on AKT1 protein that can impact the activity of the AKT1 protein was investigated by using molecular docking and molecular dynamics (MD) simulations. The molecular docking results presented a relatively higher binding affinity of the mitoxantrone molecule in interaction with AKT1 than the XFE molecule. These results were validated by the MM/PBSA technique that was performed on obtained trajectories of 25ns MD simulations. The mitoxantrone molecule has an intense binding energy of - 880.536kcal/mol with AKT1 protein, while the XFE molecule shows a binding energy value of - 83.569kcal/mol. Our findings from molecular dynamics simulations indicated that both molecules have favorite interactions with AKT1 protein. Other analyses, such as RMSF and hydrogen binding on trajectories obtained from MD simulations, indicated that the mitoxantrone molecule could be a relatively potent inhibitor for AKT1. Based on the results of this study and the structure of mitoxantrone, it is expected to be a good candidate for cancer treatment as a (PI3K)/Akt/mTOR inhibitor.

Abstract 3153: Prediction of metabolites from cancer-associated fibroblasts that can have profound effects on cancer cells

Abstract Cancer-associated fibroblasts (CAFs) are the most abundant component of tumor microenvironment (TME) that is an ensemble of non-cancerous cells surrounding cancer cells. In particular, CAFs are known to have tumor-assisting functions, which may promote active growth and survival of the interacting cancer cells through their metabolic reprogramming despite being in a harsh environment (e.g., hypoxia or low pH). In this regard, CAF-derived metabolites are expect to have profound effects on metabolism of the cancer cells. To identify novel CAF-derived metabolites that may heavily influence the cancer cells, CAF-specific genome-scale metabolic models (GEMs) were reconstructed and simulated. GEM is a computational model that can predict a metabolic state in a cell-specific manner by integrating omics data (e.g., RNA-seq data). In this study, a total of six GEMs were reconstructed by integrating RNA-seq data from one normal fibroblast, originating from pancreas, and five CAFs associated with breast, colon, skin, lung, and pancreatic tumors. Simulation of the GEMs predicted 20 metabolites that appeared to be heavily used by all the five CAF GEMs in comparison with the one normal fibroblast GEM. Such metabolites were expected to be significantly associated with establishment of TME that is favorable for the cancer cells’ growth. Some of these metabolites are associated with pyrimidine metabolism, fatty acid metabolism, and primary bile acid biosynthesis. Upon experimental validation, the predicted metabolites are expected to help better understand CAFs and TME in terms of metabolites, and also provide clues for identifying effective drug targets for cancer treatment. Citation Format: Hae Deok Jung, Yoo Jin Sung, Jae Yong Ryu, Hyun Uk Kim. Prediction of metabolites from cancer-associated fibroblasts that can have profound effects on cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3153.

Targeting Apoptotic Pathway of Cancer Cells with Phytochemicals and Plant-Based Nanomaterials.

Apoptosis is the elimination of functionally non-essential, neoplastic, and infected cells via the mitochondrial pathway or death receptor pathway. The process of apoptosis is highly regulated through membrane channels and apoptogenic proteins. Apoptosis maintains cellular balance within the human body through cell cycle progression. Loss of apoptosis control prolongs cancer cell survival and allows the accumulation of mutations that can promote angiogenesis, promote cell proliferation, disrupt differentiation, and increase invasiveness during tumor progression. The apoptotic pathway has been extensively studied as a potential drug target in cancer treatment. However, the off-target activities of drugs and negative implications have been a matter of concern over the years. Phytochemicals (PCs) have been studied for their efficacy in various cancer cell lines individually and synergistically. The development of nanoparticles (NPs) through green synthesis has added a new dimension to the advancement of plant-based nanomaterials for effective cancer treatment. This review provides a detailed insight into the fundamental molecular pathways of programmed cell death and highlights the role of PCs along with the existing drugs and plant-based NPs in treating cancer by targeting its programmed cell death (PCD) network.

Recent Advancements in the Design of Nanodelivery Systems of siRNA for Cancer Therapy.

RNA interference (RNAi) has increased the possibility of restoring RNA drug targets for cancer treatment. Small interfering RNA (siRNA) is a promising therapeutic RNAi tool that targets the defective gene by inhibiting its mRNA expression and stopping its translation. However, siRNAs have flaws like poor intracellular trafficking, RNase degradation, rapid kidney filtration, off-targeting, and toxicity, which limit their therapeutic efficiency. Nanocarriers (NCs) have been designed to overcome such flaws and increase antitumor activity. Combining siRNA and anticancer drugs can give synergistic effects in cancer cells, making them a significant gene-modification tool in cancer therapy. Our discussion of NCs-mediated siRNA delivery in this review includes their mechanism, limitations, and advantages in comparison with naked siRNA delivery. We will also discuss organic NCs (polymers and lipids) and inorganic NCs (quantum dots, carbon nanotubes, and gold) that have been reported for extensive delivery of therapeutic siRNA to tumor sites. Finally, we will conclude by discussing the studies based on organic and inorganic NCs-mediated siRNA drug delivery systems conducted in the years 2020 and 2021.

Discovery of 2-Methyl-2-(4-(2-methyl-8-(1H-pyrrolo[2,3-b]pyridin-6-yl)-1H-naphtho[1,2-d]imidazol-1-yl)phenyl)propanenitrile as a Novel PI3K/mTOR Inhibitor with Enhanced Antitumor Efficacy In Vitro and In Vivo.

PI3K/Akt/mTOR signaling pathway is a validated drug target for cancer treatment that plays a critical role in controlling tumor growth, proliferation, and apoptosis. However, no FDA-approved PI3K/mTOR dual inhibitor exists. Thus, a candidate with a better curative effect and lower toxicity is still urgently needed. Herein, we design, synthesize, and evaluate compounds belonging to a novel series of 2-methyl-1H-imidazo[4,5-c]quinoline scaffold derivatives as PI3K/mTOR dual inhibitors. Among them, compound 8o was identified as a novel candidate with excellent kinase selectivity. It manifested remarkable antiproliferative activities against SW620 and HeLa cells. Western blot and immunohistochemical analysis results proved that 8o could regulate the PI3K/AKT/mTOR signaling pathway by inhibiting the phosphorylation of AKT and S6 proteins. Additionally, 8o presented a favorable pharmacokinetic property (oral bioavailability of 76.8%) and significant antitumor efficacy in vivo without obvious toxicity. Collectively, these results indicated that 8o is a promising agent for cancer treatment and merits further development.

Abstract 3370: Fatty acid desaturase 1 (FADS1) is associated with cancer survival via altering cell growth and can be a novel drug target for cancer treatment

Abstract Polyunsaturated fatty acid (PUFA) plays an important role in cancer biology. Fatty acid desaturase1 (FADS1) is a key rate-limiting enzyme in omega-3 and omega-6 PUFA metabolism. We aim to investigate the association between FADS1 and survival outcomes among cancer patients. Univariable and multivariable Cox regression models were performed for FADS1 mRNA expression levels on disease-free survival (DFS) and overall survival (OS) using The Cancer Genome Atlas (TCGA) public datasets. We also investigated the pathways associated with altered FADS1 expression in the TCGA dataset. A FADS1 inhibitor (D5D-IN-326) was used to study the proliferation in vitro in 6 different cancer cell lines. FADS1-shRNA knockdown system was introduced in the renal cell carcinoma (RCC) 786-O cell line to further study the signaling pathway associated with proliferation. Transcriptomic, metabolomic, and lipidomic profiles of stable cell lines were studied. Our analysis showed that among all cancer patients (N=11,490), those with a higher FADS1 mRNA expression in their tumor have a significantly worse DFS and OS (p.value <0.001 and 0.01, respectively). This association was particularly significant in RCC subtypes. Pathway analysis demonstrated that cell cycle and DNA replication were significantly enriched as the top pathways associated with FADS1 expression. Administration of FADS1 inhibitor compound 326 decreased cell proliferation in cell lines of several major cancers in a dose-dependent manner. The proliferation result was confirmed in RCC FADS1 knockdown model. RNA-seq analysis of FADS1 knockdown stable cells also showed downregulation of DNA biosynthesis, cell division, and cell proliferation. In our metabolomic study, 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) was significantly reduced in FADS1 knockdown (KD) cells which may be attributed to altered purine metabolism (p-value < 0.001). As we expected, the total omega-6 and omega-3 fatty acids were lower in KD cells. We further identified lower P38 phosphorylation in KD cells which is consistent with lower cell proliferation. These results suggest that FADS1 overexpression is prognostic to lower survival in cancer patients via changing cancer metabolism, which is potentially mediated via altering cell proliferation through MAPK pathway. This study has a potential to discover a new mechanism of cancer biology and develop a novel precision treatment for cancer patients. The detailed causal relationship is warranted for continued investigation. Citation Format: Gioia Heravi, Hyejeong Jang, Seongho Kim, Wanqing Liu. Fatty acid desaturase 1 (FADS1) is associated with cancer survival via altering cell growth and can be a novel drug target for cancer treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3370.

Recent Advances in Combretastatin A-4 Inspired Inhibitors of Tubulin Polymerization: An Update.

Cancer is one of the leading causes of fatality and mortality worldwide. Investigations on developing therapeutic strategies for cancer are supported throughout the world. The massive achievements in molecular sciences involving biochemistry, molecular chemistry, medicine, and pharmacy, and high throughput techniques such as genomics and proteomics have helped create new potential drug targets for cancer treatment. Microtubules are very attractive targets for cancer therapy because of the crucial roles they play in cell division. In recent years, lots of efforts have been put into the identification of new microtubule-targeting agents (MTAs) in anticancer therapy. Combretastatin A-4 (CA-4) is a natural compound that binds to microtubules' colchicine binding site and inhibits microtubule polymerization. Due to CA-4's structural simplicity, many analogs have been synthesized. This article summarises the new molecule development efforts to reach CA-4 analogues by pharmacophore group modifications, which have been reported since 2015.

Expression of Hv1 proton channels in myeloid-derived suppressor cells (MDSC) and its potential role in T cell regulation

Myeloid-derived suppressor cells (MDSC) are a heterogeneous cell population with high immunosuppressive activity that proliferates in infections, inflammation, and tumor microenvironments. In tumors, MDSC exert immunosuppression mainly by producing reactive oxygen species (ROS), a process triggered by the NADPH oxidase 2 (NOX2) activity. NOX2 is functionally coupled with the Hv1 proton channel in certain immune cells to support sustained free-radical production. However, a functional expression of the Hv1 channel in MDSC has not yet been reported. Here, we demonstrate that mouse MDSC express functional Hv1 proton channel by immunofluorescence microscopy, flow cytometry, and Western blot, besides performing a biophysical characterization of its macroscopic currents via patch-clamp technique. Our results show that the immunosuppression by MDSC is conditional to their ability to decrease the proton concentration elevated by the NOX2 activity, rendering Hv1 a potential drug target for cancer treatment.

Calcium Channel Blockers: The Effect of Glutathione S‐Transferase Enzyme Activity and Molecular Docking Studies

AbstractRecently, as a drug target in cancer treatment, the superfamily of glutathione S‐transferase (GSTs, EC 2.5.1.18) have been invited considerable interest by scientists. In particular, as they are overexpressed in many human cancer cell lines, GSTs can catalyze the conjugation of the cellular nucleophile glutathione (GSH) with a wide range of electrophilic carcinogens toxins and drugs, meanwhile producing oxidative stress. For this purpose, the GST was purified by GSH‐agarose affinity chromatography, and some calcium channel blockers (CCBs), such as amlodipine, cinnarizine, isradipine, nifedipine, and nilvadipine, were assessed for their inhibitory actions against GST. The CCBs demonstrated micromolar levels inhibitory activity towards GST (KIs spanning within the 98.84±0.53 μM–502.70±2.53 μM range). The best GST inhibitory activity was observed for the isradipine. Additionally, molecular docking study was performed for competitive inhibitor nilvadipine on GST to describe the possible interaction with the active site to confirm the inhibitory activity.

Importance of the biomass formulation for cancer metabolic modeling and drug prediction

SummaryGenome-scale metabolic reconstructions include all known biochemical reactions occurring in a cell. A typical application is the prediction of potential drug targets for cancer treatment. The precision of these predictions relies on the definition of the objective function. Generally, the biomass reaction is used to illustrate the growth capacity of a cancer cell. Today, seven human biomass reactions can be identified in published metabolic models. The impact of these differences on the metabolic model predictions has not been explored in detail. We explored this impact on cancer metabolic model predictions and showed that the metabolite composition and the associated coefficients had a large impact on the growth rate prediction accuracy, whereas gene essentiality predictions were mainly affected by the metabolite composition. Our results demonstrate the importance of defining a consensus biomass reaction compatible with most human models, which would contribute to ensuring the reproducibility and consistency of the results.

The deubiquitinase USP10 restores PTEN activity and inhibits non–small cell lung cancer cell proliferation

The phosphatase and tensin homolog deleted on chromosome 10 (PTEN) protein is a key player in tumorigenesis of non–small cell lung cancer (NSCLC) and was recently found to be inactivated by tripartite motif containing 25 (TRIM25)–mediated K63-linked polyubiquitination. However, the deubiquitinase (Dub) coordinate TRIM25 in PTEN ubiquitination is still elusive. In the present study, we found that this K63-linked polyubiquitination could be ablated by the ubiquitin-specific protease 10 (USP10) in a screen against a panel of Dubs. We found using coimmununoprecipitation/immunoblotting that USP10 interacted with PTEN and reduced the K63-linked polyubiquitination of PTEN mediated by TRIM25 in NSCLC cells. Moreover, USP10, but not its inactive C424A deubiquitinating mutant or other Dubs, abolished PTEN from K63-linked polyubiquitination mediated by TRIM25. In contrast to TRIM25, USP10 restored PTEN phosphatase activity and reduced the production of the secondary messenger phosphatidylinositol-3,4,5-trisphosphate, thereby inhibiting AKT/mammalian target of rapamycin progrowth signaling transduction in NSCLC cells. Moreover, USP10 was downregulated in NSCLC cell lines and primary tissues, whereas TRIM25 was upregulated. Consistent with its molecular activity, re-expression of USP10 suppressed NSCLC cell proliferation and migration, whereas knockout of USP10 promoted NSCLC cell proliferation and migration. In conclusion, the present study demonstrates that USP10 coordinates TRIM25 to modulate PTEN activity. Specifically, USP10 activates PTEN by preventing its K63-linked polyubiquitination mediated by TRIM25 and suppresses the AKT/mammalian target of rapamycin signaling pathway, thereby inhibiting NSCLC proliferation, indicating that it may be a potential drug target for cancer treatment.

Conformational Clamping by a Membrane Ligand Activates the EphA2 Receptor

The EphA2 receptor is a promising drug target for cancer treatment, since EphA2 activation can inhibit metastasis and tumor progression. It has been recently described that the TYPE7 peptide activates EphA2 using a novel mechanism that involves binding to the single transmembrane domain of the receptor. TYPE7 is a conditional transmembrane (TM) ligand, which only inserts into membranes at neutral pH in the presence of the TM region of EphA2. However, how membrane interactions can activate EphA2 is not known. We systematically altered the sequence of TYPE7 to identify the binding motif used to activate EphA2. With the resulting six peptides, we performed biophysical and cell migration assays that identified a new potent peptide variant. We also performed a mutational screen that determined the helical interface that mediates dimerization of the TM domain of EphA2 in cells. These results, together with molecular dynamic simulations, allowed to elucidate the molecular mechanism that TYPE7 uses to activate EphA2, where the membrane peptide acts as a molecular clamp that wraps around the TM dimer of the receptor. We propose that this binding mode stabilizes the active conformation of EphA2. Our data, additionally, provide clues into the properties that TM ligands need to have in order to achieve activation of membrane receptors.

A novel Apigenin derivative suppresses renal cell carcinoma via directly inhibiting wild-type and mutant MET

MET, the receptor of hepatocyte growth factor (HGF), is a driving factor in renal cell carcinoma (RCC) and also a proven drug target for cancer treatment. To improve the activity and to investigate the mechanisms of action of Apigenin (APG), novel derivatives of APG with improved properties were synthesized and their activities against Caki-1 human renal cancer cell line were evaluated. It was found that compound 15e exhibited excellent potency against the growth of multiple RCC cell lines including Caki-1, Caki-2 and ACHN and is superior to APG and Crizotinib. Subsequent investigations demonstrated that compound 15e can inhibit Caki-1 cell proliferation, migration and invasion. Mechanistically, 15e directly targeted the MET kinase domain, decreased its auto-phosphorylation at Y1234/Y1235 and inhibited its kinase activity and downstream signaling. Importantly, 15e had inhibitory activity against mutant MET V1238I and Y1248H which were resistant to approved MET inhibitors Cabozantinib, Crizotinib or Capmatinib. In vivo tumor graft study confirmed that 15e repressed RCC growth through inhibition of MET activation. These results indicate that compound 15e has the potential to be developed as a treatment for RCC, and especially against drug-resistant MET mutations.

Prognostic value of plasma hPG80 (circulating progastrin), alone or in combination with Alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma.

3033 Background: Alpha-fetoprotein (AFP) is the most widely used biomarker for hepatocellular carcinoma (HCC) prognosis since it is expressed in the advanced stages of the disease. Consequently, AFP is not useful in establishing a prognosis for patients with a tumor in the early stages of the disease. hPG80 (circulating progastrin), a new drug target for cancer treatment which plays a pivotal role in tumorigenesis, is present in the blood of multiple types of cancers at early stages including HCC. The purpose of this study was to evaluate the prognostic value of plasma hPG80 in patients with HCC, in combination or not with AFP. Methods: A total of 168 HCC patients (BCLC from 0 to D) managed with local or systemic treatments, (“Liverpool” biobank) were enrolled prospectively and analyzed retrospectively. hPG80 was quantified using DxPG80 Lab kit (ECS-Progastrin) and AFP was quantified using Cobas E411 in the blood of HCC patients. An optimal cutoff value of hPG80 was identified at 4.5 pM by calculating the minimal p-value based on the log-rank method. For AFP, a cutoff of 100 ng/mL was used as for liver transplantation (Notarpaolo, 2016). The prognostic impact of hPG80 and AFP levels on patient survival was assessed using Kaplan-Meier curves and log-rank tests. Results: The median overall survival (OS) of the full cohort is 20.9 months. HCC patients with high hPG80 levels (hPG80+: >4.5 pM, 105/168) had significantly lower median OS compared to patients with low hPG80 levels (hPG80-: <4.5 pM, 63/168) (12.4 months versus undefined respectively, p < 0.0001). Patients with high AFP (AFP+: >100 ng/mL, 69/165) had significantly lower median OS compared to patients with low AFP (AFP: <100 ng/mL 96/165) (7.2 months versus undefined, p < 0.0001). To improve the stratification, the patients were further categorized into four groups: hPG80-/AFP- (n = 42), hPG80+/AFP- (n = 54), hPG80-/AFP+ (n = 21) and hPG80+/AFP+ (n = 48). In the AFP- group, hPG80+ patients exhibited a significantly worse prognosis than those with hPG80- (26.3 months versus undefined, p=0.0087). Similarly, in the AFP+ group, patients with hPG80+ had a significantly worse survival compared to hPG80- patients (5.7 months versus 13.4 months, p = 0.0391). Finally, we evaluated the median OS of AFP+ patients according to BCLC staging. Interestingly, in the group BCLC 0 to B, hPG80+ had a significantly worse prognosis than those with hPG80- (15.8 months versus 40.25 months, p=0.0317). Conclusions: Our findings show that hPG80 could serve as a new prognostic biomarker in HCC. Used in combination with AFP, it improves the stratification of the patients in good and worst prognosis, especially for those patients with negative AFP and early-stage HCC.

PPARs and Tumor Microenvironment: The Emerging Roles of the Metabolic Master Regulators in Tumor Stromal-Epithelial Crosstalk and Carcinogenesis.

Simple SummaryThe roles of peroxisome proliferator-activated receptors (PPARs) in carcinogenesis are increasingly appreciated. With the growing interest in tumor stromal-epithelial crosstalk, we aim to provide an up-to-date overview of the implications of PPARs in the tumor microenvironment. In the tumor stromal cells, the nuclear receptors exhibit critical, but functionally diverse activities, rendering it hard to ascribe either an exclusive pro- or anti-tumorigenic role for different PPAR isotypes. Based on the existing evidence, we also highlight the knowledge gaps and future prospects of targeting PPARs in the tumor microenvironment. Essentially, a PPAR-based anticancer approach holds a great deal of untapped potential, but its success relies on innovative strategies for cell-specific or tumor microenvironment-triggered drug delivery systems.Peroxisome proliferator-activated receptors (PPARs) have been extensively studied for more than three decades. Consisting of three isotypes, PPARα, γ, and β/δ, these nuclear receptors are regarded as the master metabolic regulators which govern many aspects of the body energy homeostasis and cell fate. Their roles in malignancy are also increasingly recognized. With the growing interest in crosstalk between tumor stroma and epithelium, this review aims to highlight the current knowledge on the implications of PPARs in the tumor microenvironment. PPARγ plays a crucial role in the metabolic reprogramming of cancer-associated fibroblasts and adipocytes, coercing the two stromal cells to become substrate donors for cancer growth. Fibroblast PPARβ/δ can modify the risk of tumor initiation and cancer susceptibility. In endothelial cells, PPARβ/δ and PPARα are pro- and anti-angiogenic, respectively. Although the angiogenic role of PPARγ remains ambiguous, it is a crucial regulator in autocrine and paracrine signaling of cancer-associated fibroblasts and tumor-associated macrophages/immune cells. Of note, angiopoietin-like 4 (ANGPTL4), a secretory protein encoded by a target gene of PPARs, triggers critical oncogenic processes such as inflammatory signaling, extracellular matrix derangement, anoikis resistance and metastasis, making it a potential drug target for cancer treatment. To conclude, PPARs in the tumor microenvironment exhibit oncogenic activities which are highly controversial and dependent on many factors such as stromal cell types, cancer types, and oncogenesis stages. Thus, the success of PPAR-based anticancer treatment potentially relies on innovative strategies to modulate PPAR activity in a cell type-specific manner.

Identification and analysis of genes associated with lung adenocarcinoma by integrated bioinformatics methods.

Lung adenocarcinoma (LUAD) is one of the most common forms of lung cancer, with a very high mortality rate. Although the treatments available for LUAD have become more effective in recent years, significant improvement is still needed. Advances in sequencing technologies and bioinformatics analysis have enabled new approaches to be developed for identifying drug targets. In this work we utilized data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to identify hub genes related to LUAD through Weighted Gene Correlation Network Analysis (WGCNA) and other bioinformatics methods, with the goal of identifying new drug targets for cancer treatment.