Tumor Volume In Mice Research Articles

Dual targeting of Mg/N doped-carbon quantum dots with folic and hyaluronic acid for targeted drug delivery and cell imaging

Mg/N doped-carbon quantum dots (CQDs) with dual drug targeting and cell imaging properties was synthesized. Mg/N doped-CQDs synthesized by a hydrothermal method. Operating pyrolysis parameters such as temperature, time, and pH were optimized to achieve CQDs with high quantum yield (QY). This CQD applied in cellular imaging. For the first time, dual active targeting of Mg/N doped CQDs performed using folic acid and hyaluronic acid (CQD-FA-HA). Then, epirubicin (EPI) loaded on this nanocarrier as the final complex (CQD-FA-HA-EPI). Cytotoxicity analysis, cellular uptake, and cell photography performed for the complex on three cell lines, including 4T1, MCF-7, and CHO. In vivo studies were performed in BALB/c inbred female mice models bearing breast cancer. Characterization results showed the successful formation of Mg/N doped-CQDs with a high QY of 89.44%. In vitro drug release approved pH dependency of synthesized nanocarrier with a controlled release behavior. Cytotoxicity tests and cellular uptake results demonstrated increased toxicity and absorption into 4T1 and MCF-7 cell lines for targeted nanoparticles compared to free drug. In cell imaging, an increase in the entry of the complex into 4T1 and MCF-7 cells compared to free drug, confirmed the proper function of the synthesized complex. In vivo results indicated that the tumor volume of mice receiving CQD-FA-HA-EPI was the lowest among other studied groups, along with the lowest damage to the liver, spleen, and heart according to the histopathological analysis. Finally, CQD-FA-HA proposed as a novel platform with tumor targeting, drug carrier, and photoluminescence properties.

A phase 1 trial of umbilical cord blood–derived tumor-reactive PD-L1+ natural killer cells engineered to express soluble IL-15 (TRACK-NK) in patients with non–small-cell lung cancer (NSCLC) refractory to PD-1/PD-L1 inhibitors.

TPS2665 Background: The mechanism by which patients with PD-L1 – tumors can respond favorably to anti-PD-L1 monoclonal antibody therapy has been shown to involve the PD-L1+ NK cell in the tumor microenvironment (TME). PD-L1 is induced on the cell surface when NK cells recognize tumors, leading to enhanced NK-cell function without exhaustion. The PD-L1 + NK cell is capable of trafficking to the lung TME, recognizing and killing tumor cells as well as inducing activation of endogenous T cells via potent cytokine secretion, all without the expression of a chimeric antigen receptor (CAR). TRACK-NK cells are PD-L1+ NK cells derived from cord blood (CB) and engineered to express soluble IL-15 (sIL-15). The TRACK-NK cells express high levels of tumor reactive receptors that recognize "tumor stress" including DNAM-1, NKp30 and NKG2D. TRACK-NK cells induced significant reduction in tumor volume in mice injected with human A549 NSCLC cells compared to mock-transduced NK cells, or NK cells expressing sIL-15 (sIL-15-NK) but without ex vivo activation. Survival was also prolonged in the H460 NSCLC model compared to vehicle alone. We hypothesized that TRACK-NK cells can be safely administered, and will provide meaningful clinical benefit in patients with NSCLC. Methods: NCT05334329 is a first-in-human, phase 1 dose finding study of allogeneic, off-the shelf frozen and thawed TRACK-NK cells in patients with NSCLC progressing on treatment with PD-1/PD-L1 inhibitors. TRACK-NK cells (CB NK cells engineered with retroviral transduction to express sIL-15 and ex vivo expanded and activated to express PD-L1) are administered in four weekly intravenous infusions per cycle following lymphocyte depletion (Fludarabine/Cyclophosphamide x 3 days) in the outpatient setting. Patients not experiencing disease progression (per RECIST v1.1) following cycle 1 may receive a second cycle. Primary endpoints include: 1. Toxicity/adverse events as per NCI-CTCA version 5.0, and ASTCT Consensus Grading for Cytokine Release Syndrome (CRS) and Neurotoxicity; and 2. Magnitude/duration of TRACK-NK cell persistence in peripheral blood via droplet digital PCR. We are employing a Utility Based-Bayesian Optimal Interval design to direct optimal dose determination. Dose limiting toxicity is defined as any ≥ grade 3 non-hematologic toxicity; any grade 3 CRS that does not resolve to ≤ grade 1 within 7 days; any ≥ grade 4 CRS; any ≥ grade 2 neurotoxicity that does not resolve to grade 1 within 72 hours; any ≥ grade 3 autoimmune, hypersensitivity, and acute graft-versus-host reactions. Correlatives also include cytokines levels, quantity and activation of CD4 and CD8 cells in peripheral blood, and NK and T cell trafficking and TME changes in paired tumor biopsies using quantitative immunofluorescence and gene expression analysis. Clinical trial information: NCT05334329 .

Cell penetrating peptides: Highlighting points in cancer therapy.

Cell-penetrating peptides (CPPs), first identified in HIV a few decades ago, deserved great attention in the last two decades; especially to support the penetration of anticancer drug means. In the drug delivery discipline, they have been involved in various approaches from mixing with hydrophobic drugs to the use of genetically conjugated proteins. The early classification as cationic and amphipathic CPPs has been extended to a few more classes such as hydrophobic and cyclic CPPs so far. Developing potential sequences utilized almost all methods of modern science: choosing high-efficiency peptides from natural protein sequences, sequence-based comparison, amino acid substitution, obtaining chemical and/or genetic conjugations, in silico approaches, in vitro analysis, animal experiments, etc. The bottleneck effect in this discipline reveals the complications that modern science faces in drug delivery research. Most CPP-based drug delivery systems (DDSs) efficiently inhibited tumor volume and weight in mice, but only in rare cases reduced their levels and continued further processes. The integration of chemical synthesis into the development of CPPs made a significant contribution and even reached the clinical stage as a diagnostic tool. But constrained efforts still face serious problems in overcoming biobarriers to reach further achievements. In this work, we reviewed the roles of CPPs in anticancer drug delivery, focusing on their amino acid composition and sequences. As the most suitable point, we relied on significant changes in tumor volume in mice resulting from CPPs. We provide a review of individual CPPs and/or their derivatives in a separate subsection.

Facilitation of Temozolomide Resistance of Glioblastoma by Long Noncoding RNA DLK1-35

Background: Long noncoding RNAs played critical roles in glioblastoma development. Objectives: This study aimed to examine the impacts of lncRNA DLK1-35 on glioblastoma cells and mice. Methods: Methyl Thiazolyl Tetrazolium (MTT) was applied for examining the viabilities of U87 and U251 cells, as well as IC50 values of temozolomide (TMZ). LncRNA DLK1-35 expressions were detected using RT-qPCR. Proliferation and apoptosis of TMZ-resistant U251 (U251 TR) cells were evaluated using colony formation and flow cytometry, respectively. Western blot was applied to analyze O6-methylguanine-DNA methyltransferase (MGMT) protein expressions. The xenograft model was used for detecting the weight and volume of tumors in mice. Results: TMZ treatment suppressed the viabilities of glioblastoma cells dose-dependently. Moreover, TMZ-resistant glioblastoma cells had higher IC50 values. lncRNA DLK1-35 was upregulated in TMZ-resistant cells while the suppression of lncRNA DLK1-35 caused low proliferation and a higher apoptosis rate. Moreover, MGMT was also inhibited by lncRNA DLK1-35 downregulation. Additionally, the weight and volume of tumors in mice were also inhibited with the knockdown of lncRNA DLK1-35. Conclusion: Knockdown of lncRNA DLK1-35 inhibited MGMT to decrease the TMZ resistance in vitro and in vivo in glioblastoma.

Abstract 1433: Targeting the mitotic kinase NEK2 potentiates CDK4/6 inhibitor efficacy in breast cancer

Abstract Mitotic kinases are essential for cell cycle progression, and several mitotic kinase inhibitors have been developed to target the uncontrolled proliferation of cancer cells. However, despite their efficacy in preclinical models, mitotic kinase inhibitors are generally considered poor therapeutic targets due to their off target effects in other rapidly proliferating tissues and subsequent toxicity. However, preliminary data in our lab shows that relative to non-transformed cells, triple negative breast cancer (TNBC) cells are selectively sensitive to inhibition of the mitotic kinase NEK2 compared to other mitotic kinase inhibitors. Therefore, we hypothesize that NEK2 is a targetable vulnerability of TNBC. To begin to understand the role of NEK2 in TNBC, we performed RNA-sequencing following NEK2 silencing and found increased expression of genes important at the G1/S cell cycle transition. This transition is largely controlled by the RB-E2F signaling axis, where CDK4 and CDK6 are key players. CDK4/6 inhibitors have been FDA approved for the treatment of a subset of breast cancers. We investigated the impact of silencing NEK2 and treating with CDK4/6 inhibitors and found that loss of NEK2 sensitizes TNBC cells to CDK4/6 inhibition. Mechanistically, this combination causes a significant increase in genomic instability phenotypes. We tested the combination of NEK2 inhibition and CDK4/6 inhibition in vivo, and found that the combination significantly decreases mouse tumor volume without inducing toxicity. Mechanistically, the combination altered mitotic spindle genes in vivo, indicating an increase in genomic instability. Together, these data suggest that that NEK2 is a viable therapeutic target that could be used in combination with FDA approved CDK4/6 inhibitors for the treatment of aggressive breast cancers such as TNBC. Citation Format: Jessica R. Bobbitt, Leslie Cuellar-Vite, Benjamin Bryson, Kristen Weber-Bonk, Ruth Keri. Targeting the mitotic kinase NEK2 potentiates CDK4/6 inhibitor efficacy in breast cancer [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 1433.

MP20-07 NPTX1 IS A NOVEL SERUM BIOMARKER INDUCED BY ARPI TREATED PROSTATE CANCER AND ENRICHED IN LINEAGE PLASTICITY WITH BRN2 HIGH

You have accessJournal of UrologyCME1 Apr 2023MP20-07 NPTX1 IS A NOVEL SERUM BIOMARKER INDUCED BY ARPI TREATED PROSTATE CANCER AND ENRICHED IN LINEAGE PLASTICITY WITH BRN2 HIGH Takeshi Namekawa, Joshua Scurll, Maxim Kobelev, Dwaipayan Ganguli, Olena Sivak, Martin Gleave, Vancouver, Canada, Eva Corey, Colm Morrissey, and Amina Zoubeidi Takeshi NamekawaTakeshi Namekawa More articles by this author , Joshua ScurllJoshua Scurll More articles by this author , Maxim KobelevMaxim Kobelev More articles by this author , Dwaipayan GanguliDwaipayan Ganguli More articles by this author , Olena SivakOlena Sivak More articles by this author , Martin GleaveMartin Gleave More articles by this author , Vancouver Vancouver More articles by this author , Canada Canada More articles by this author , Eva CoreyEva Corey More articles by this author , Colm MorrisseyColm Morrissey More articles by this author , and Amina ZoubeidiAmina Zoubeidi More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003245.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Androgen receptor (AR) pathway inhibition is the first-line therapy for metastatic prostate cancer (PCa). However, tumors invariably develop resistance to ARPI. We previously identified a transcription factor Brain-2 (BRN2), encoded by the gene POU3F2, as a master driver of NEPC which is an aggressive form of AR-independent phenotypes treatment resistant PCa. In this study, we identified a surrogate biomarker of BRN2 activity to monitor the progression of treatment resistant PCa. METHODS: To identify BRN2 serum marker, we analyzed BRN2 ChIPseq data from 2 NEPC and 2 neural-type lung squamous cell carcinoma (nLUSC) cell lines with a focus on shared BRN2 peaks. Annotated peaks from enhancers (EnhancerAtlas) and promoters (genes within 20kb) were selected for secreted proteins and were ranked by correlation with BRN2 expression in PCa patients. Validation of protein expression was performed by western blot. R-PLEX Assay kit was used to measure concentrations in culture media from NCI-H660 cells and in serum from mice with NCI-H660 xenograft or PDX tumors and from human PCa patients. RESULTS: We identified 51 candidate serum biomarkers for BRN2 activity (or expression?) using the methods described above. These biomarkers were ranked based on their expression and correlation with BRN2 expression in PCa patient samples across four public data sets. NPTX1 was identified as the top biomarker of BRN2 expression. We further validated association of NPTX1 and BRN2 expression by BRN2 overexpression in MR42D and 22Rv1, indicating that BRN2 positively regulates NPTX1 expression. Next, we showed that NPTX1 is present in culture media from NCI-H660 cells, and that NPTX1 serum concentration positively correlated with tumor volume in mice with NCI-H660 xenografts and with NPTX1 mRNA expression in LuCaP PDX models. Importantly, evaluation of patients’ samples showed that NPTX1 levels were high in 26/35 CRPC serum samples vs levels in serum of healthy volunteers, and that the NPTX1 serum levels were highest in 4/6 patients with high POU3F2 tumor expression. CONCLUSIONS: We identified and validated NPTX1 as a serum marker that can be readily detected in serum from NEPC patients with BRN2high tumours. Source of Funding: U.S. Army Medical Research and Development Command(W81XWH1810689) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e277 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takeshi Namekawa More articles by this author Joshua Scurll More articles by this author Maxim Kobelev More articles by this author Dwaipayan Ganguli More articles by this author Olena Sivak More articles by this author Martin Gleave More articles by this author Vancouver More articles by this author Canada More articles by this author Eva Corey More articles by this author Colm Morrissey More articles by this author Amina Zoubeidi More articles by this author Expand All Advertisement PDF downloadLoading ...

Adoption of Ultrasound-guided Polymer Nanocarriers in Tumor Chemoradiotherapy and Oxidation Treatment.

The research was aimed at discussing the effectiveness of ultrasound-guided polymer nanocarriers in the clinical treatment of tumors by chemoradiotherapy and oxidation treatment. Twenty female Balb/cAnN (BALB/C) mice were selected as the research objects in the experiment. These mice were set up as tumor-bearing mice, and then ultrasound-guided polymers with different doses, including polyethylene glycol-poly 2-bromoethyl methacrylate (PEG-PBEMA) (Micelle group), free small molecules called l-ascorbyl palmitate (PA) (PA group), PA-micelle micellar particles (PA-Micelle group) prepared in the research, and phosphate buffer solution (PBS) (PBS group) were adopted. Besides, the growth of mice was recorded and compared after each operation. Meanwhile, different concentrations of PA-Micelle micellar particles and free small molecules of PA were added to the breast cancer cells of mice, and the concentration changes of glutathione (GSH) were detected to test the oxidation treatment ability of this method. According to the results of the experiment, the tumor volume of mice in the PA-Micelle group prepared in the research was the smallest followed by the PA group, and the tumor volume of mice in the Micelle group was the third smallest. The mice in the PBS group had the largest tumors among mice in all four groups. In oxidation treatment, the GSH concentration of mice in the PA-Micelle group was the lowest, while the GSH concentration of mice in the PA group was almost unchanged. The results of this experiment proved that the therapeutic effect of polymer nanocarriers in tumor chemotherapy and oxidation treatment was more significant than in traditional drug treatment.

Nodakenin Induces ROS-Dependent Apoptotic Cell Death and ER Stress in Radioresistant Breast Cancer

Angelica gigas exerts powerful anti-tumor and anti-cancer effects in various cancer cell types. However, there have been few studies regarding the anti-cancer effect of nodakenin, a bioactive compound of Angelica gigas, in vivo and in vitro on breast cancers. I found that nodakenin, in a concentration-dependent manner, inhibits breast cancer cell viability and decreases the tumor volume in mice. Additionally, nodakenin induces caspase-3-dependent apoptosis in breast cancer cells; however, the combination of Z-VAD-FMK and nodakenin suppresses the caspase-3-dependent apoptotic cell death. Furthermore, nodakenin mediates apoptotic cell death via the PERK-mediated signaling pathway and calcium (Ca2+) release, and nodakenin combined with thapsigargin induces synergistic cell death by inhibiting sarco/endoplasmic reticulum (ER) Ca2+-ATPase. However, knockdown of PERK or CHOP inhibits Ca2+ generation and caspase-dependent apoptosis in nodakenin-treated breast cancer cells. Nodakenin induces ROS and Ca2+ generation, ER stress, and apoptotic cell death; however, the knockdown of Nox4 inhibits ROS generation and ER stress- and caspase-dependent apoptotic cell death. In addition, nodakenin combined with radiation overcomes radioresistance in radioresistant breast cancer cells by suppressing epithelial-mesenchymal transition phenotypes, including the decrease in E-cadherin and the increase in N-cadherin and vimentin. Therefore, these findings indicate that nodakenin may be a novel therapeutic strategy for breast cancers.

Compound K: A systematic review of its anticancer properties and probable mechanisms.

Panax ginseng is a common natural product, which is well-known to have a wide range of pharmacological activities in cancer. Its metabolite, compound K (CK), has been reported to have anticancer activity. We aimed to systematically review the literature for evidence of anticancer effects of CK. We conducted a systematic search in eight databases. We included all in vitro and in vivo studies investigating the anticancer effects of CK with no restrictions. Quality assessment was applied by ToxRTool. Fifty-four articles were included in our study. The purity of CK in our included studies was at least 95%. The in vitro studies reported that CK had a potential anticancer activity on several cell lines including human lung cancer cell lines (A549, PC-9), nasopharyngeal carcinoma cell line (Hk-1), liver cancer cell line (BEL 7402), and pediatric acute myeloid leukemia cell lines (Kasumi-1, MV4-11). The in vivo studies reported a significant decrease in tumor volume in mice treated with CK. CK is a potential supplementary treatment in cancer chemotherapies. The safety and further clinical trials of CK should be explored for future drug development.

Antitumor effect of selenium-rich Brazil nuts and selenomethionine dietary supplementation on pre-existing 4T1 mammary tumor growth in mice.

Selenium (Se) is an essential micronutrient known to play an important role in the antioxidant system that can potentially influence tumor growth. We aimed to investigate the effects of dietary Se supplementation after detection of 4T1 mammary tumor growth in BALB/c mice. Thirty female mice received subcutaneous inoculation of 4T1 cells. After five days, all animals presenting palpable tumors were randomly assigned to three groups: a control group (Se-control) receiving a diet with adequate Se (0.15 mg/kg) and two other groups that received Se-supplemented diets (1.4 mg/kg of total Se) with either Brazilian nuts (Se-Nuts) or selenomethionine (SeMet). Data were assessed by either One or Two-way ANOVA followed by Tukey's HSD or Bonferroni's post hoc tests, respectively. Both Se-supplemented diets reduced tumor volume from the thirteenth day of feeding compared with the Se-adequate (control) diet (p < 0.05). The SeMet group presented a higher Se blood concentration (p < 0.05) than the Se-control group, with the Se-Nuts group presenting intermediate values. Selenoprotein P gene expression in the liver was higher in the Se-Nuts group than in the Se-control group (p < 0.05), while the SeMet group presented intermediate expression. Dietary Se supplementation, starting after detection of 4T1 palpable lesions, reduced tumor volume in mice.

Porphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by activating the neutrophil chemotaxis in the tumour microenvironment.

We aimed to determine the significance of Porphyromonas gingivalis (P. gingivalis) in promoting tumour progression in the tumour microenvironment (TME) of oral squamous cell carcinoma (OSCC). The Gene Expression Omnibus (GEO) was used to screen out the differentially expressed genes from the two datasets of GEO138206 and GSE87539. Immunohistochemistry and immunofluorescence analysis of samples, cell biological behaviour experiments, and tumour-bearing animal experiments were used to verify the results in vivo and in vitro. The mechanism was revealed at the molecular level, and rescue experiments were carried out by using inhibitors and lentiviruses. CXCL2 was selected by bioinformatics analysis and was found to be related to a poor prognosis in OSCC patients. Samples with P. gingivalis infection in the TME of OSCC had the strongest cell invasion and proliferation and the largest tumour volume in tumour-bearing animal experiments and exhibited JAK1/STAT3 signalling pathway activation and epithelial-mesenchymal transition (EMT). The expression of P. gingivalis, CXCL2 and TANs were independent risk factors for poor prognosis in OSCC patients. A CXCL2/CXCR2 signalling axis inhibitor significantly decreased the invasion and proliferation ability of cells and the tumour volume in mice. When lentivirus was used to block the CXCL2/CXCR2 signalling axis, the activity of the JAK1/STAT3 signalling pathway was decreased, and the phenotype of EMT was reversed. Porphyromonas gingivalis promotes OSCC progression by recruiting TANs via activation of the CXCL2/CXCR2 axis in the TME of OSCC.

Root Bark Extract of Oroxylum indicum Vent. Inhibits Solid and Ascites Tumors and Prevents the Development of DMBA-Induced Skin Papilloma Formation

Oroxylum indicum is a traditionally used plant in Ayurvedic and folk medicines. The plant is useful for the management of gastrointestinal diseases as well as skin diseases. In the present study, we analyzed the antitumor potential of O. indicum in Dalton's lymphoma ascites tumor cells (DLA) and Ehrlich ascites carcinoma (EAC)-induced solid and ascites tumors. Further, the potential of O. indicum extract (OIM) on skin papilloma induction by dimethyl benz(a) anthracene (DMBA) and croton oil was evaluated. The chemical composition of the extract was analyzed using UPLC-Q-TOF-MS. The predominant compounds present in the extract were demethoxycentaureidin 7-O-rutinoside, isorhamnetin-3-O-rutinoside, baicalein-7-O-glucuronide, 5,6,7-trihydroxyflavone, 3-Hydroxy-3',4',5'-trimethoxyflavone, 5,7-dihydroxy-3-(4-methoxyphenyl) chromen-4-one, and 4'-Hydroxy-5,7-dimethoxyflavanone. Treatment with high-dose OIM enhanced the percentage of survival in ascites tumor-bearing mice by 34.97%. Likewise, high and low doses of OIM reduced the tumor volume in mice by 61.84% and 54.21%, respectively. Further, the skin papilloma formation was brought down by the administration of low- and high-dose groups of OIM (by 67.51% and 75.63%). Overall, the study concludes that the Oroxylum indicum root bark extract is a potentially active antitumor and anticancer agent.

Effect of Guizhi Fuling Capsule on Apoptosis of Myeloma Cells Through Mitochondrial Apoptosis Pathway.

To observe the effects of Guizhi Fuling Capsule (GZFLC) on myeloma cells and explore the mechanisms. MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell experiments were divided into negative (10%, 20% and 40%) groups, GZFLC (10%, 20%, and 40%) groups and a control group. Cell counting kit-8 (CCK-8) assays and flow cytometry were used to detect the viability and apoptosis levels of myeloma cells. The effects on mitochondria were examined by reactive oxygen specie (ROS) and tetrechloro-tetraethylbenzimidazol carbocyanine iodide (JC-1) assays. Western blot was used to detect the expression of B cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), cleaved caspase-3, -9, cytochrome C (Cytc) and apoptotic protease-activating factor 1 (Apaf-1). RPMI 8226 cells (2 × 107) were subcutaneously inoculated into 48 nude mice to study the in vivo antitumor effects of GZFLC. The mice were randomly divided into four groups using a completely randomized design, the high-, medium-, or low-dose GZFLC (840, 420, or 210 mg/kg per day, respectively) or an equal volume of distilled water, administered daily for 15 days. The tumor volume changes in and survival times of the mice in the GZFLC-administered groups and a control group were observed. Cytc and Apaf-1 expression levels were detected by immunohistochemistry. GZFLC drug serum decreased the viability and increased the apoptosis of myeloam cells (P<0.05). In addition, this drug increased the ROS levels and decreased the mitochondrial membrane potential (P<0.01). Western blot showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups, whereas the expression levels of cleaved caspase-3, -9, Cytc and Apaf-1 were increased (all P<0.01). Over time, the myeloma tumor volumes of the mice in the GZFLC-administered groups decreased, and survival time of the mice in the GZFLC-administered groups were longer than that of the mice in the control group. Immunohistochemical analysis of tumor tissues from the mice in the GZFLC-administered groups revealed that the Cytc and Apaf-1 expression levels were increased (P<0.05). GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway and significantly reduced the tumor volumes in mice with myeloma, which prolonged the survival times of the mice.

Antitumor Effect and Pharmacological Mechanism of Paclitaxel-loaded Silk Fibroin Nanomaterials on H22 Subcutaneous Tumor Model of Mouse Hepatoma

This work was developed to analyze the adoption of paclitaxel-loaded silk fibroin nanoparticles (SFNPs) in subcutaneous transplanted tumor model of mouse hepatoma to explore its anti-tumor effect. Twenty-five specific pathogen-free (SPF) mice were selected to construct a subcutaneous tumor model of liver cancer, and they were randomly rolled into control group, group A, group B, group C, and group D, with five mice in each group. The silk fibroin was mixed with an organic solvent to prepare a suspension, and the SFNPs were prepared through centrifugation, ultrasound, and other methods. The paclitaxel-loaded SFNPs were prepared by mixing paclitaxel and silk fibroin aqueous solution, centrifuging, washing, and dispersing. Then, the five groups of mice were intervened by different dosage regimens to analyze the changes of various indicators. As a result, the prepared nanoparticles had uniform particle size, uniform distribution, no adhesion, and the average particle size was less than 500 nm. The tumor volume of mice in groups C and D on the 7th, 9th, and 13th days of administration were dramatically smaller than those in the control group, group A, and group B (P [ 0.05). And the tumor volume (154.49 � 9.65 mm3) of mice in group D on the 13th day of administration was dramatically smaller than that in group C (167.79 � 9.72 mm3) (P [ 0.05). The tumor mass (0.89 � 0.14 g, 0.54 � 0.13 g, and 0.46 � 0.11 g) of mice in groups B, C, and D was dramatically smaller than that in the control group and group A (1.23 � 0.12 g, 1.24 � 0.11 g) (P [ 0.05), and that of group D was dramatically smaller than groups B and C (P [ 0.05). The apoptosis rates of tumor cells in groups C and D (46.38%, 48.23%) were greatly superior to those in the control group (16.7%), group A (21.33%), and group B (35.6%) (P [ 0.05), and that of group D was greatly superior to that in group C (P [ 0.05). In summary, paclitaxel-loaded SFNPs can effectively improve the targeting effect and bioavailability of drugs in the treatment of liver cancer, thereby improving the efficacy, and had a good application prospect.

Abstract B014: Metabolic reprogramming in high-grade sarcomas: repurposing anti-cholesterol agents as a novel therapeutic strategy

Abstract Introduction and Objective: Undifferentiated pleomorphic sarcoma (UPS) and leiomyosarcoma (LMS) are treated with surgery, radiation, and/or chemotherapy. Current drug therapies, such as doxorubicin, have limited response rates and severe toxicities. We performed a drug screen with over 3000 compounds on four patient-derived UPS cell lines and uncovered sensitivity to simvastatin, which inhibits the rate limiting enzyme in the mevalonate pathway. Statins are well tolerated drugs used to lower cholesterol in patients with hypercholesterolemia. The mevalonate pathway can be exploited by cancer through metabolic reprogramming and phosphoinositide 3-kinase/mammalian target of rapamycin (PI3K/mTOR) signaling to enhance proliferation. Thus, the goals of our study are to define the mechanism(s) responsible for simvastatin sensitivity and determine the efficacy when combined with doxorubicin in vivo. Hypothesis: Dysregulation of the mevalonate pathway and/or PI3K/mTOR pathway will render certain high-grade sarcomas sensitive to simvastatin. Methods: We confirmed simvastatin sensitivity in UPS, as well as in LMS cell lines, with dose-response curves. We investigated the mechanisms whereby simvastatin inhibits UPS and LMS viability with immunoblotting and flow cytometry. Proliferation was quantified using a cell proliferation dye. To assess in vivo efficacy and toxicity of simvastatin we performed a murine study with an established xenograft UPS model from our lab. We are currently assessing the efficacy of this novel combination in an LMS xenograft. Results: We found decreased phosphorylated AKT (pAKT ser473) and increased rates of active caspase-3 in most sensitive UPS cell lines with simvastatin (1uM) treatment. This indicates downregulation of PI3K/mTOR signaling and an increase in apoptosis, respectively. Importantly, these effects were rescued with the addition of mevalonate (200uM), an effector immediately downstream of the rate limiting enzyme that statins inhibit, indicating an on-target effect. In four sensitive LMS cell lines treated with simvastatin (1.5uM or 2uM) a decrease in pAKT ser473 is not consistently seen, revealing that downregulation of PI3K/mTOR signalling may not be a universal mechanism. Furthermore, in some UPS and LMS cell lines there was a significant reduction in proliferation with simvastatin + doxorubicin (0.5uM) treatment (p&amp;lt;0.05). Our in vivo study demonstrated that simvastatin (50mg/kg) in combination with doxorubicin (1.2mg/kg) reduced the volume of UPS tumors in mice. Furthermore, mice treated with combination therapy had higher concentrations of simvastatin and doxorubicin within the tumor than those treated with either single agent. However, this study was not designed to look at pharmacokinetics (PK), so to confirm these results a PK study is underway in an LMS xenograft. Conclusion: UPS and LMS are sensitive to simvastatin, suggesting these sarcomas rely on metabolic reprogramming to sustain viability. Thus, simvastatin may be a novel therapy for certain sarcoma patients. Citation Format: Jen Dorsey, Yael Babichev, Rosemarie E. Venier, Richard Marcellus, Rima Al-awar, Linda Z. Penn, Albiruni A. Razak, Brendan C. Dickson, Eric Chen, Irene L. Andrulis, Jay Wunder, Rebecca A. Gladdy. Metabolic reprogramming in high-grade sarcomas: repurposing anti-cholesterol agents as a novel therapeutic strategy [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B014.

Tumor imaging and photothermal therapy in second near infrared window: A systematic review and meta-analysis.

BackgroundSecond near-infrared window (NIR-II, 1000-1700 nm) technology for tumor imaging and photothermal therapy (PTT) is an innovative method for tumor diagnosis and treatment. The NIR-II probe can specifically identify tumor cells, and effectively convert light energy into heat energy under the irradiation of NIR laser, thus achieving the integration of non-invasive tumor diagnosis and treatment. In the present study, we conducted a systematic review and meta-analysis of preclinical investigations to corroborate the efficacy and safety of photothermal therapy.MethodsRelevant preclinical data were retrieved by searching PubMed, Web of Science, CNKI, WANFANG and VIP information databases. And the acquired data were analyzed by RevMan Version 5.3 software.ResultsAccording to the inclusion criteria, forty-two articles relating to NIR-II tumor imaging and PTT were recruited for further in-depth analysis. The NIR-II photoacoustic and fluorescence imaging could quickly and accurately identify tumor in mice, manifesting higher signal intensity on tumor site than that of normal tissue. After PTT, the tumor volume of mice decreased miraculously [RR=8.49, 95%CI (4.64, 15.55), P<0.00001], and even disappeared completely [RR=7.01, 95%CI (3.04, 16.13), P<0.00001] with no potential risk of affecting the blood routine.ConclusionsPTT guided by NIR-II imaging can effectively diagnose the tumor lesion and eliminate it with the advantages of non-invasive and higher biosafety.

Silibinin exhibits anti-tumor effects in a breast cancer stem cell model by targeting stemness and induction of differentiation and apoptosis.

Introduction: Malignant breast cancer (BC) frequently contains a rare population of cells called cancer stem cells which underlie tumor relapse and metastasis, and targeting these cells may improve treatment options and outcomes for patients with BC. The aim of the present study was to determine the effect of silibinin on the self-renewal capacity, tumorgenicity, and metastatic potential of mammospheres. Methods: The effect of silibinin on viability and proliferation of MCF-7, MDA-MB-231 mammospheres, and MDA-MB-468 cell aggregation was determined after 72-120 hours of treatment. Colony and sphere formation ability, and the expression of stemness, differentiation, and epithelial-mesenchymal-transition (EMT)-associated genes were assessed by reverse transcription-quantitative polymerase chain reaction (qRT-PCR) in mammospheres treated with an IC50 dose of silibinin. Additionally, the antitumor capacity of silibinin was assessed in vivo, in mice. Results: The results of the present study showed that silibinin decreased the viability of all mammospheres derived from MCF-7, MDA-MB-231, and MDA-MB-468 cell aggregation in a dose-dependent manner. Colony and sphere-forming ability, as well as the expression of genes associated with EMT were reduced in mammospheres treated with silibinin. Additionally, the expression of genes associated with stemness and metastasis was also decreased and the expression of genes associated with differentiation were increased. Intra-tumoral injection of 2 mg/kg silibinin decreased tumor volumes in mice by 2.8 fold. Conclusion: The present study demonstrated that silibinin may have exerted its anti-tumor effects in BC by targeting the BC stem cells, reducing the tumorgenicity and metastasis. Therefore, silibinin may be a potential adjuvant for treatment of BC.

Anticancer effect of Indanone-based thiazolyl hydrazone derivative on p53 mutant colorectal cancer cell lines: An in vitro and in vivo study.

Colorectal cancer is a major health problem, and it is the third most diagnosed cancer in the United States. The current treatment for colorectal cancer includes irinotecan, a topoisomerase I inhibitor, and other targeted drugs, such as bevacizumab and regorafenib. The low response rates and incidence of high toxicity caused by these drugs instigated an evaluation of the anticancer efficacy of a series of 13 thiazolyl hydrazone derivatives of 1-indanone, and four compounds among them show favorable anticancer activity against some of the tested colorectal cancer cell lines with IC50 values ranging from 0.41 ± 0.19 to 6.85 ± 1.44 μM. It is noteworthy that one of the indanone-based thiazolyl hydrazone (ITH) derivatives, N-Indan-1-ylidene-N’-(4-Biphenyl-4-yl-thiazol-2-yl)-hydrazine (ITH-6), has a better cytotoxicity profile against p53 mutant colorectal cancer cells HT-29, COLO 205, and KM 12 than a p53 wild-type colorectal cancer cell line, such as HCT 116. Mechanistic studies show that ITH-6 arrests these three cancer cell lines in the G2/M phase and induces apoptosis. It also causes a rise in the reactive oxygen species level with a remarkable decrease in the glutathione (GSH) level. Moreover, ITH-6 inhibits the expression of NF-κB p65 and Bcl-2, which proves its cytotoxic action. In addition, ITH-6 significantly decreased tumor size, growth rate, and tumor volume in mice bearing HT-29 and KM 12 tumor xenografts. Moreover, CRISPR/Cas9 was applied to establish an NF-κB p65 gene knockout HT-29 cell line model to validate the target of ITH-6. Overall, the results suggest that ITH-6 could be a potential anticancer drug candidate for p53 mutant colorectal cancers.

Chloroquine Enhances BIIB021-induced Apoptosis in Chronic Myeloid Leukemia Cells Bearing T315I Mutation

To explore the combined pro-apoptosis effect of HSP90 inhibitor BIIB021 and chloroquine (CQ) in chronic myeloid leukemia (CML) cells bearing T315I mutation and its mechanism. The p210-T315I cells were divided into 4 groups by different treatment: control, BIIB021, CQ, and BIIB021 + CQ. After treated with BIIB021 or/and CQ for 24 hours, Annexin V/PI binding assay was used to detect apoptosis rates of CML cells. DAPI staining was used to observe nuclear fragmentation, and Western blot was used to detect the expression of caspase 3, PARP (apoptosis related proteins) and p62, LC3-I/II (autophagy related proteins). P210-T315I cells were inoculated subcutaneously into mice and CML mouse models were established. The mice in treatment groups were injected with BIIB021 and/or CQ while mice in control group were treated with PBS and normal saline. The tumor volume of mice was measured every 4 days, and protein level of cleaved-caspase 3 and LC3-II in tumor tissue were detected by immunohistochemistry. The results showed that BIIB021 induced apoptosis of CML cells in a dose-dependent manner ( r=0.91). CQ could enhance the apoptosis-inducing effect of BIIB021. Flow cytometry analysis results showed that the apoptosis rate of p210-T315I cells in combination group was higher than that in BIIB021 or CQ only group (P<0.05). DAPI staining showed nuclear fragmentation in combination group could be observed more obviously. Western blot analysis showed that BIIB021 could induce LC3-I to convert to LC3-II and decrease p62 protein levels (P<0.05). Moreover, the combination group had higher expression of LC3-II, p62 (P<0.05), activated PARP and activated caspase 3 than BIIB021 only group (P<0.05). Besides, experiment in vivo showed the mean tumor volume in co-treatment group was lower than that in single drug group (P<0.01). Immunohistochemistry of tumor tissue also showed the protein level of cleaved-caspase 3 and LC3-II in combined group was higher than that in BIIB021 only group. HSP90 inhibitor BIIB021 induced significant apoptosis of CML cells bearing T315I both in vivo and in vitro. CQ can enhance this effect probably by autophagy inhibition.

Small molecules to regulate the GH/IGF1 axis by inhibiting the growth hormone receptor synthesis.

Growth hormone (GH) and insulin‐like growth factor‐1 (IGF1) play an important role in mammalian development, cell proliferation and lifespan. Especially in cases of tumor growth there is an urgent need to control the GH/IGF1 axis. In this study we screened a 38,480-compound library, and in two consecutive rounds of analogues selection, we identified active lead compounds based on the following criteria: inhibition the GH receptor (GHR) activity and its downstream effectors Jak2 and STAT5, and inhibition of growth of breast and colon cancer cells. The most active small molecule (BM001) inhibited both the GH/IGF1 axis and cell proliferation with an IC50 of 10‐30 nM of human cancer cells. BM001 depleted GHR in human lymphoblasts. In preclinical xenografted experiments, BM001 showed a strong decrease in tumor volume in mice transplanted with MDA‐MB‐231 breast cancer cells. Mechanistically, the drug acts on the synthesis of the GHR. Our findings open the possibility to inhibit the GH/IGF1 axis with a small molecule.