Antitumor Potential Research Articles

An engineered α1β1 integrin-mediated FcγRI signaling component to control enhanced CAR macrophage activation and phagocytosis

Treatment of solid tumors remains difficult, and therefore there has been increased focus on chimeric antigen receptor macrophages (CAR-M) to challenge solid tumors. However, CAR domain design of of adoptive cell therapy, which leads to differences in antitumor activity and triggered antitumor potential, remains poorly understood for macrophages. We developed an α1β1 integrin-mediated Fc-gamma receptor I (FcγRI) signaling component for CAR-M specific activation and its antitumor potential. We evaluated CAR-M effects with α1β1 integrin-mediated FcγRI signaling (ACT CAR-M) on the activation and antitumor phagocytic response of macrophages in vitro. Subcutaneous tumor model in BALB/c mice and carcinomatosis model in immunodeficient mice were used to test the antitumor effect of ACT CAR-M compared with CD3ζ CAR-M. The α1β1 integrin-mediated FcγRI signaling engagement of CAR-M was associated with enhanced macrophage activation and specific phagocytosis in primary human macrophages, and significantly improved tumor control and survival in multiple cancer models when compared to CD3ζ CAR-M. RNA-sequencing suggested that α1β1 integrin-mediated FcγRI engagement increased antitumor immunity by enhancing pro-inflammatory M1 phenotype-associated pathways, such as Toll-like receptor signaling, tumor necrosis factor signaling, and IL-17 signaling. α1β1 integrin-mediated FcγRI signaling engagement markedly enhanced antitumor effects of CAR-M immunotherapy, which is proposed as an advanced engineering CAR domain material to expand the clinical application of CAR-M.

Pirfenidone promotes cell cycle arrest and apoptosis of triple‑negative breast cancer cells by suppressing Hedgehog/GLI1 signaling.

Breast cancer is a common malignant tumor in women and triple-negative breast cancer (TNBC) is the most challenging type of breast cancer with poor prognosis. We aimed to elucidate the effects of pirfenidone, a FDA-approved oral anti-fibrotic drug which has recently shown antitumor potential, in the progression of TNBC and the underlying mechanisms. After TNBC cells were treated with pirfenidone, cell viability was evaluated using CCK-8 assay. The EDU staining was applied for reflecting the ability of cell proliferation. Additionally, cell cycle distribution and apoptotic rate of TNBC cells exposed to pirfenidone were determined by flow cytometry. The levels of proteins associated with cell cycle, apoptosis and Hedgehog/gliomaassociated oncogene homolog (GLI)1 signaling was examined through western blot. Then, GLI1 was upregulated to analyze the proliferation, cell cycle and apoptosis of TNBC cells in the presence of pirfenidone to reveal the regulatory mechanism. Pirfenidone dose-dependently decreased the viability and proliferation of MDA-MB-231 and HCC-1937 cells. Besides, the distribution of TNBC cells in G0/G1 phase was significantly elevated by pirfenidone, accompanied by downregulated levels of CyclinD1, CDK4 and CDK6. Simultaneously, pirfenidone caused remarkably increased apoptotic MDA-MB-231 and HCC-1937 cells, coupled with downregulated BCL2 expression as well as upregulated Bax, cleaved caspase3 and cleaved PARP expression. Expression of proteins related to Hedgehog/gliomaassociated oncogene homolog (GLI)1 signaling was tested through western blot. Particularly, GLI1 and PTCH1 levels were dose-dependently inhibited after pirfenidone exposure. Interestingly, GLI1 overexpression attenuated the influences of pirfenidone on the proliferation, cell cycle and apoptosis of TNBC cells. Collectively, pirfenidone arrests the cell cycle and promotes apoptosis of TNBC cells by suppressing Hedgehog/GLI1 signaling.

Quantitative Characterization of the Effect of Biogenic silver-based Nanoparticles on Breast Cancer Cells by High Content Analysis

Background: Among the various types of cancer, breast cancer is the most incident among women. Due to the resistance to antitumor treatments, alternative treatments have been sought, such as metallic nanoparticles. Objective: This study aimed to evaluate the antitumor potential and cytotoxicity induction mechanisms of green synthesized AgCl-NPs and Ag/AgCl-NPs. Methods: The antitumor potential of nanoparticles was evaluated in breast cancer BT-474 and MDAMB- 436 cell lines treated with 0-40 μg/mL AgCl-NPs or 0-12.5 μg/mL Ag/AgCl-NPs through imagebased high content analysis method. Normal human retinal pigment epithelial 1 (RPE-1) cells were used for comparison. Results: The growth rate of the RPE-1 cells treated with nanoparticles was insignificantly affected, and no significant changes in cell viability were observed. In these cells, the nanoparticle treatments did not induce lysosomal damage, changes in ROS production, or reduction in the mitochondrial membrane potential. The level of BT-474 and MDA-MB-436 cell proliferation was markedly decreased, and cell viability was reduced by 64.19 and 46.19% after treatment with AgCl-NPs and reduced by 98.36 and 82.29% after treatment with Ag/AgCl-NPs. The cells also showed a significant increase in ROS production and loss of mitochondrial membrane potential, which culminated in an increase in the percentage of apoptotic cells. BT-474 cells also presented lysosomal damage when treated with the highest concentrations of both nanoparticle types and actin polymerization was observed after exposure to Ag/AgCl-NPs. Conclusions: Together, the results showed overall cytotoxic effects of both AgCl-NPs and Ag/AgCl- NPs towards breast cancer cells with negligible effects against healthy cells, which suggests their promising anticancer and biomedical applications.

Ethnomedicinal uses, Pharmacological activity, and Analytical aspects of Galuteolin: A Natural Flavonoid of Medicinal Importance

Background: Nature is the best source of all raw materials that we need for growth, development, and better health. Flavonoids are an important class of phytochemicals found to be present in plants and their derived byproducts. Lonicera japonica and its active phytochemicals possess antiviral, anti-endotoxin, anti-pyretic, and blood fat-reducing properties in medicine. Moreover, Galuteolin is a natural flavonoid that has anti-oxidant, anti-tumor, anti-infective, and antiinflammatory potential. Methods: In the present review article, we have collected all scientific information on galuteolin from different scientific databases and analyzed it to know the biological importance and therapeutic benefit of galuteolin in medicine against human disorders. Scientific data on galuteolin were collected from numerous literature databases, such as Google, Google Scholar, PubMed, Scopus and Science Direct. Detailed pharmacological data on galuteolin were collected from these scientific sources and analyzed in the present work to know the health-beneficial aspects of galuteolin in medicine. In addition, scientific data on analytical techniques of galuteolin were also collected and analyzed in the present work to know their importance in the separation, isolation, and identification of galuteolin in different plant materials. Results: The present work signified the biological importance of galuteolin in medicine and other allied health sectors. Pharmacological data analysis revealed the health-beneficial aspects of galuteolin on rheumatoid arthritis, cerebral ischemia, carcinoma, COVID-19, Atherosclerosis, and melanin biosynthesis. Further, intestinal absorption and pharmacokinetic parameters of galuteolin were also discussed in the present work. Analytical data signified the importance of newer and modern analytical techniques in medicine for the qualitative and quantitative analysis of galuteolin in different biological samples. Conclusion: Scientific data analysis of galuteolin revealed its biological importance and therapeutic benefit in medicine.

Dichlorophosphoryl Isocyanate: Synthesis, Chemical Reactions, and Biological Activity of its Derivatives

Abstract: Dichlorophosphoryl isocyanate (DCPI) is the most basic and easy phosphoric isocyanate substrate that introduces organic moieties. Synthesized since 1954, the DCPI has a high reactivity toward primary and secondary alkyl, alcohols, phenols, thiols, and amines via the nucleophilic addition reaction on the carbon atom of the isocyanate group. In addition to their synthesis, the resulting products undergo nucleophilic substitutions of the chlorine atoms. Their reactions with nucleophilic and bi-nucleophile reagents yield acyclic and P-heterocyclic compounds, respectively. The resulting compounds have different potential antibacterial, antifungal, and antitumor activities.

Cellulose nanocrystals/cellulose nanofibrils-combined astaxanthin nanoemulsion for reinforcement of targeted tumor delivery of gastric cancer cells

Nanoemulsion based nanomaterial (NE) was carried out in the present study to evaluate the efficacy and its antitumor potential of the gastric cancer cells. NE was prepared with astaxanthin/alpha-tocopherol- cellulose nanocrystals/cellulose nanofibrils based nanoemulsions for gastric cancer treatment. The cytotoxic potential was tested against cancer cells and evaluated in terms of its cell proliferation, migration, and cellular uptake by the standard methods. NE was examined for its synergetic effect with photodynamic therapy (PDT) in a xenograft mouse model. The results confirmed the synergetic effect of PDT and NEs in the in vivo animal model. The regulated expression of proteins manifested the reduced toxicity and inhibition of cell proliferation and migration. The antitumor study showed that NE inhibited the growth of human colon cancer in vivo. Immunohistological analysis confirmed the regulation of PI3K/AKT signaling pathway. The present study demonstrates that NEs can enhance anti-cancer effect against human gastric cancer through the immunomodulatory signaling pathway.

Antitumor potentials of onco-microbial in Chinese patients with pancreatic cancer

Recent studies have revealed that intratumoral microbiota is implicated in pancreatic cancer (PC), yet the spectra of intratumoral microbiota and their relationship with PC in Chinese patients remained to be clarified. In this study, tumor and paired paracancerous tissue from 53 patients were profiled by bacterial 16S rRNA gene sequencing. Both α- and β-diversity displayed significant differences between tumors and adjacent tissues, with higher diversity in tumors. Three bacteria phyla (Proteobacteria, Firmicutes, and Actinobacteria) were prevalent in both cancers and adjacent normal tissues. A high prevalence of Pseudomonas has been identified in the PC tumor microenvironment and was associated with prolonged overall survival. Furthermore, the results of in vitro experiments suggested that Pseudomonas fluorescens (P. fluorescens) could inhibit the proliferation and induce apoptosis of pancreatic cancer cells. These findings revealed distinctive microbial features of the PC tumors and normal tissues in Chinese populations and exhibited the antitumor potential of P. fluorescens in PC.

Synthesis, characterization, DNA, BSA, antimicrobial, antioxidant and molecular docking studies of gadolinium(III) based complexes

The present study reports the synthesis of two new gadolinium-metal-based complexes with mixed ligands, [Gd(L1)2(py)(H2O)3] (C1) and [Gd (L2)2(phen)(H2O)2] (C2), where L1 represents 3-(4 cyanophenyl) acrylate and L2 as 3-(4 fluorophenyl) acrylate. The complexes were characterized by various analytical techniques including FT-IR, PXRD, TGA, DSC, SEM and EDX to determine structural elucidation, binding mode, crystallinity, thermal stability, morphology and elemental composition, respectively. The synthesized complexes were tested for DNA (salmon sperm) and BSA interaction studies to infer their antitumor potential. The binding constants obtained for C1 and C2 with SS DNA were 3.03 × 104 and 3.2 × 104 while with BSA observed as 1.59 × 105 and 1.22 × 105, respectively. In vitro antimicrobial potential of both complexes was determined against various bacterial and fungal strains. Experimental results showed complex C2 is more active than C1 against all selected bacterial strains and showed higher antibacterial potential against E. coli with a zone of inhibition (ZOI) of 35 mm. The antibacterial potential of C2 was further investigated by calculating the MIC (IC50) value. MIC of complex C2 was assessed using visual inspection (turbidity) and spectrophotometric assay at 600 nm. The findings of the spectrophotometric approach have greater sensitivity than the standard disc diffusion assays. The molecular docking studies were conducted to validate further and compare the findings of experimental results which exhibited that complex C2 has an excellent binding capacity with the target protein (5CDQ) with free binding energy −12.23 kcal/mol. Antioxidant potentials of the complexes were assessed using phosphomolybdenum assay, which demonstrated that complex C2 exhibits higher antioxidant activity than complex C1.

Cytotoxic, Antioxidant, and Anti-Genotoxic Properties of Combretastatin A4 in Human Peripheral Blood Mononuclear Cells: A Comprehensive In Vitro Study

Despite significant advances in drug discovery and the promising antitumor potential of combretastatin A4 (CA-4), which selectively targets rapidly dividing cancer cells, CA-4’s effects on non-dividing human cells, such as peripheral blood mononuclear cells (PBMCs), remain unclear. The aim of this study is to evaluate the in vitro bioactivity of CA-4 in human PBMCs, focusing on its antigenotoxic and antioxidant properties, while comparing its cytotoxic potency against PBMCs, cancer cell lines (JAR and HeLa), and the normal trophoblast cell line HTR-8/SVneo. Cell viability and metabolic activity were evaluated using the MTT assay. ROS production in PBMCs was measured using the H2DCFDA assay, and DNA damage was assessed using the Comet assay. CA-4 showed cytotoxicity in PBMCs and HTR-8/SVneo cells at concentrations above 200 µM, while cancer cells, JAR and HeLa, showed cytotoxicity at 100 µM and 1 µM, respectively. CA-4 also reduced ROS levels in PBMCs under oxidative stress and showed antioxidant effects at concentrations from 1 to 200 µM. In addition, CA-4 showed antigenotoxic effects against H2O2-induced DNA damage in PBMCs at concentrations of up to 1 µM. CA-4 exhibited lower cytotoxicity in human PBMCs compared to cancer cells, inhibited ROS production, and showed antioxidant and antigenotoxic properties, providing insight into its potential therapeutic efficacy and safety.

Study on the Chemical Composition and Anti-Tumor Mechanisms of Clausena lansium Fruit By-Products: Based on LC-MS, Network Pharmacology Analysis, and Protein Target Validation

Clausena lansium (Lour.) Skeels, commonly known as Wampee, are valued for their edible and medicinal qualities, yet their pericarp and seeds are often discarded, resulting in wasted resources. This study investigates the anti-tumor potential of these by-products, focusing on their chemical composition and underlying mechanisms of action. A combination of metabolomics, network pharmacology, molecular docking, and experimental validation was employed in our study. Cytotoxicity screening demonstrated that the pericarp extract exhibited notable anti-tumor effects against MDA-MB-231 breast cancer cells, while the seed extract showed no similar activity. Chemical profiling identified 122 compounds in the pericarp and seeds, with only 26.23% overlap, suggesting that distinct compounds may drive the pericarp’s anti-tumor activity. Network pharmacology and molecular docking analyses identified PTGER3, DRD2, and ADORA2A as key targets, with several alkaloids, flavonoids, coumarins, and sesquiterpenes exhibiting strong binding affinities to these proteins. Western blot analysis further validated that the pericarp extract upregulated DRD2 and downregulated ADORA2A, indicating a possible mechanism for its anticancer effects. These findings suggest that Wampee pericarp holds promise as a source of active compounds with therapeutic potential for breast cancer, with implications for its use in the food and pharmaceutical industries.

Assessing Novel Antibody-Based Therapies in Reconstructive 3D Cell Models of the Tumor Microenvironment.

Targeted, combinatorial, and immunomodulatory therapies, such as antibody-drug conjugates (ADCs) and immunomodulatory antibodies (Abs), are powerful weapons against tumor cells and immune cells within the tumor microenvironment (TME). Therefore, the evaluation of such therapies should be conducted in pre-clinical models able to recapitulate the complex cellular and molecular crosstalk of the TME. To build-in critical hallmarks of the TME, a breast cancer heterotypic 3D cell model (3D-3) is devised using a microencapsulation strategy with an inert biomaterial (alginate) and agitation-based cultures. Both stromal and immune components are added to multicellular tumor spheroids, therefore fostering cancer-associated fibroblasts (CAFs) and tumor-associated macrophages (TAMs) immunomodulatory interactions. The potential of the methodology to assess Ab-based therapies is then addressed by employing a series of anti-HER2-based ADCs. ADCs induced tumor-cell specific cytotoxicity toward HER2+ breast cancer spheroids while sparing HER2-negative CAFs. In addition, an immunomodulatory blocking Ab against colony-stimulating factor 1 receptor (CSF1R) decreases the expression of immunosuppressive and anti-inflammatory markers in TAMs, like what is previously observed upon in vivo α-CSF1R administration. Collectively, the human TME-based 3D-3 cell model is a suitable tool to evaluate the anti-tumor and immunomodulatory potential of novel antibody-based therapies directed against TME targets, such as cancer cells and macrophages.

Withaferin-A induced vimentin S56 phosphorylation dissociates NEDD9 signaling loop to regress progressive metastatic melanoma into lung adenocarcinoma

Metastasis is complex and insidious type of disease involves multiple signaling nexus, which have implications in understanding disease pathogenesis. Treatment failure for metastatic cancer is frequently high due to aggressive adaptation of cancerous cells to invade to neighboring organs. Cytoskeleton intermediate filamentous protein Vimentin and scaffolding protein Neural precursor cell expressed Developmentally Down-regulated protein 9 (NEDD9) play a key role in metastatic events by regulating multiple metastatic events. Interaction between these proteins is necessary to promote metastatic progression. Withaferin A (WFA), a natural pharamacophore, known to target Vimentin to induce antitumor potential. However exact molecular mechanism still yet to be elucidated. We hypothesize, Vimentin-NEDD9 signaling nexus is necessary for metastatic progression and targeting this interwoven signaling loop with effective pharamacophore WFA halts metastatic progression of melanoma into lung. To elucidate the same, we carried out gene expression measurement through quantitative Reverses Transcription Polymerase Chain Reaction (qRT-PCR), Immunoblot and Immunohistochemistry. Assessment of interactive signaling by Co-immunoprecipitation, Immunofluorescence, Co-localization and Proximity ligation assay. Phosphorylation studies through transfection of phospho specific mutant constructs generated through site directed mutagenesis. WFA induced cellular behavioral changes by migration, invasion assays and Immunoblot analysis. The B16F10 induced mouse metastatic melanoma model to asses NEDD9-Vimentin expression and anti-metastasis induced by WFA. The results postulates, elevated levels and interaction between NEDD9-Vimentin proteins, have positive correlation in metastatic progression of melanoma into lung in both in-vitro and in-vivo condition, establishing it as therapeutic target. Pharmacologically, WFA targets this complex by extending its activity by not only inducing specific Serine 56 phosphorylation of Vimentin, also dissociates NEDD9 signaling loop to halt Epithelial-mesenchymal transition (EMT) and subsequent metastatic events. Eventually, modulation of the relevant metastatic genes E-Cadherin, N-Cadherin, SNAIL, MMP-2 & MMP-9 resulted in regression of metastatic melanoma progression to lung. The study validates WFA induced S56 phosphorylation is necessary to abrupt the NEDD9-Vimentin metastatic signaling complex to regress aggressive metastatic melanoma. The investigation emphasized more mechanistic approach of WFA. Understanding and targeting such integrative mechanical input in the tumor microenvironment will be a better therapeutic strategy to combat metastasis.

Isolation and biomimetic synthesis of acylphloroglucinol meroterpenoids as anti-breast cancer agents from Dryopteris crassirhizoma.

Two new acylphloroglucinol-nerolidol meroterpenoids (APNMs) [(±)-1 and (±)-2], with a skeleton of mixed sesquiterpene and dimeric acylphloroglucinol biosynthetic origin, were isolated from the medicinal pteridophyte Dryopteris crassirhizoma. Inspired by the proposed biosynthetic pathway, we initially completed the biomimetic syntheses of eight optically active Dryopteris APNMs (1-8) in a one-pot domino reaction. The structures of APNMs 1-8 including their absolute configurations were unambiguously established by a combination of NMR analysis, ECD calculations, and synthetic methods. Furthermore, an underlying method based on NMR data to assign the stereochemistry of the long-chain alcohol moieties in APNMs was revealed. Anti-triple negative breast cancer (anti-TNBC), antifungal and antibacterial activities of the synthetic meroterpenoids were evaluated. All tested compounds exhibited cytotoxicity against TNBC cells MDA-MB-231 with IC50 values in the range of 1.22-27.43 μM, with (-)-8 being the most potent antitumor agent (IC50: 0.48 μM; selectivity index: 32.04). This study resulted in the biomimetic synthesis of APNMs for the first time and positioned APNMs from D. crassirhizoma as a new class of promising candidates for the development of new anti-TNBC drugs.

Biochemometrics-Based Identification of Gallic Acid and Gallic Acid Galloylglucosides From Kalanchoe fedtschenkoi With Cytotoxic Effects on Cultured Melanoma Cells.

Kalanchoe Adans. (Crassulaceae) is a genus of widespread succulent plants extensively studied for their biological activities. Plants of the genus are considered a potential source of antitumor agents. This study aimed to investigate the effect of an aqueous extract and fractions of leaves of Kalanchoe fedtschenkoi R. Hamet & H. Perrier on the proliferation of melanoma cell lines employing an NMR-based biochemometric approach complemented with HPLC-DAD and UHPLC-MS/MS analyses. The n-butanol fraction (KFBuOH) from K. fedtschenkoi aqueous leaf extract, which decreased B16F10 murine melanoma cells viability by 65% at 100 μg/mL, was fractionated with RP-18 SPE and Sephadex LH-20 column chromatography. The fractions were analyzed by 1H-NMR spectroscopy and submitted to MTT cytotoxicity assays against cultured melanoma cells. Orthogonal projection to latent structures discriminant analysis (OPLS-DA) was used to correlate their 1H-NMR profile and cytotoxic activity. This strategy enabled the identification of gallic acid (1) and two gallic acid glucosides-gallic acid 4-O-(6'-O-galloyl)-glucopyranoside) (2) and gallic acid 3-O-(6'-O-galloyl)-glucopyranoside) (3)-as putative bioactive substances, which was further corroborated by subsequent assays with enriched fractions and a gallic acid standard. The fractions enriched in gallic acid (KFA) and gallic acid galloylglucosides (KFB) evidenced selective cytotoxicity towards B16F10 cells (IC50 43.0 and 56.6 μg/mL, respectively) and MV3 human melanoma cells (IC50 93.6 and 66.1 μg/mL, respectively). These results suggest a potential therapeutic use for K. fedtschenkoi in melanoma treatment. This is the first study to evidence a potential antitumor activity for gallic acid galloylglucosides.

Carnosic acid inhibits integrin expression and prevents pulmonary metastasis of melanoma.

Carnosic acid is a naturally occurring, plant-derived polyphenolic abietane diterpene with anti-tumor properties. However, its underlying mechanisms are still unclear. Therefore, we investigated the effects of carnosic acid on lung metastasis in a murine melanoma model. C57BL/6 mice were intravenously injected with B16-BL6 cells, followed by carnosic acid treatment. Lung weights were recorded, and tumor cell colonies were counted at the end of the experiment. Integrin expression was evaluated using flow cytometry and cell adhesion assays. Lung weights were significantly lower in the carnosic acid group than in the control group, indicating the suppression of metastasis. Carnosic acid suppressed α4 integrin expression in B16-BL6 cells and inhibited α4 and α9 integrin-dependent cell adhesion. Thus, our data suggest that carnosic acid prevents lung metastasis, possibly by suppressing integrin expression. Our findings support the clinical application of carnosic acid as a potential natural anti-tumor agent, offering a complementary approach to conventional therapies.

Oncolytic virus encoding 4-1BBL and IL15 enhances the efficacy of tumor-infiltrating lymphocyte adoptive therapy in HCC.

Previous studies have found that oncolytic virus (OVs) can improve the efficacy of TIL adoptive therapy in oral cancer, colon cancer, and pancreatic cancer. However, the curative effect in hepatocellular carcinoma (HCC) is still unclear. Therefore, this study aims to explore the therapeutic effect and mechanism of OVs encoding 4-1BBL and IL15 (OV-4-1BBL/IL15) combined with TIL adoptive therapy on HCC. In this study, the role and immunological mechanism of armed OVs combined with TILs were evaluated by flow cytometry and ELISA in patient-derived xenograft and syngeneic mouse tumor models. Co-culturing with TILs can up-regulate the expression of antigen-presenting cell (APC) markers on the surface of OV-infected primary HCC cells, and promote the specific activation ability and tumor-killing ability of TILs. OV-4-1BBL/IL15 combined with TIL adoptive therapy could induce tumor volume reduction and anti-tumor immune memory in patient-derived xenograft and syngeneic mouse tumor models. Furthermore, OV combined with TIL adoptive therapy can endow tumor cells with aAPC characteristics, activate T cells at the same time, and reprogram tumor macrophages into anti-tumor phenotype. OV-4-1BBL/IL15 can stimulate the anti-tumor potential of TIL therapy in HCC, and possess broad clinical application prospects.

A PSMA-targeted Tri-specific Killer Engager enhances NK cell cytotoxicity against prostate cancer.

Natural killer (NK) cell tumor infiltration is associated with good prognosis in patients with metastatic castration-resistant prostate cancer (mCRPC). NK cells recognize and kill targets by a process called natural cytotoxicity. We hypothesized that promoting an antigen-specific synapse with co-activation may enhance NK cell function in mCRPC. We describe a Tri-specific Killer Engager (TriKE) construct that engages with the activating receptor CD16 on NK cells, prostate-specific membrane antigen (PSMA) on mCRPC cells, and has an interleukin (IL)-15 moiety that is essential for NK cell survival, proliferation, and priming. We show that the PSMA TriKE specifically binds to PSMA-expressing cells and significantly enhances expansion, degranulation and cytokine production of NK cells derived from healthy donors or prostate cancer patients. Bystander killing of PSMA-negative was also achieved with PSMA TriKE treatment when co-cultured with PSMA-positive cells, suggesting potential PSMA TriKE benefit in controlling tumor antigen escape. When tested under physiologic conditions recapitulating the mCRPC tumor microenvironment (TME), NK cells treated with PSMA TriKE and prolonged exposure to hypoxia or MDSCs maintained their potent function while IL-15 treated NK cells showed greatly impaired cytotoxicity. Finally, in vivo testing of PSMA TriKE showed improved tumor control and survival of mice as compared to IL-15 and untreated control groups. In conclusion, PSMA TriKE demonstrates potential as a new therapy for advanced prostate cancer by providing additional signals to NK cells to maximize their anti-tumor potential in prostate cancer, especially in the setting of a hostile TME.

Augmenting antitumor efficacy of Th17-derived Th1 cells through IFN-γ-induced type I interferon response network via IRF7

The importance of CD4+ T cells in cancer immunotherapy has gained increasing recognition. Particularly, a specific subset of CD4+ T cells coexpressing the T helper type 1 (Th1) and Th17 markers has demonstrated remarkable antitumor potential. However, the underlying mechanisms governing the differentiation of these cells and their subsequent antitumor responses remain incompletely understood. Single-cell RNA sequencing (scRNA-seq) data reanalysis demonstrated the presence of Th171 cells within tumors. Subsequent trajectory analysis found that these Th171 cells are initially primed under Th17 conditions and then converted into IFN-γ-producing cells. Following the in vivo differentiation trajectory of Th171 cells, we successfully established in vitro Th171 cell culture. Transcriptomic profiling has unveiled a substantial resemblance between in vitro-generated Th171 cells and their tumor-infiltrating counterparts. Th171 cells exhibit more potent antitumor responses than Th1 or Th17 cells. Additionally, Th171chimeric antigen receptor T (CAR-T) cells eradicate solid tumors more efficiently. Importantly, Th171 cells display an early exhaustion phenotype while retaining stemness. Mechanistically, Th171 cells migrate faster and accumulate more in tumors in an extracellular matrix protein 1 (ECM1)-dependent manner. Furthermore, we show that IFN-γ up-regulated IRF7 to promote the type I interferon response network and ECM1 expression but decreased the exhaustion status in Th171 cells. Taken together, our findings position Th171 cells as a great candidate for improving targeted immunotherapies in solid malignancies.

Green synthesized Zingiber officinale-ZnO nanoparticles: anticancer efficacy against 3D breast cancer model.

Aim: ZnO NPs were prepared via green synthesis utilizing Zingiber Officinale.Methodology: Physical characterization and biological activity were performed against 2D, and 3D spheroids MCF-7 cell lines.Results: The NPs exhibited 188.9, 175.7 and 171.2nm size with charge of -8.2, -11.7 and -9.7mV for the 2%, 3% and 4% formulations. XRD confirmed a wurtzite hexagonal phase. FTIR spectra showed Zn-O stretching vibrations. The 2%, 3% and 4% formulations presented IC50 values of 14.7, 26.2 and 47μg/ml, respectively, with complete destruction of MCF-7 spheroids. Elevated TNF-α levels suggested an inflammatory-mediated mechanism of action.Conclusion: 2% Zingiber officinale-derived ZnO NPs showed antitumor potential against deserving further mechanistic and in vivo explorations.

Molecular Photoswitching Unlocks Glucose Oxidase for Synergistically Reinforcing Fenton Reactions for Antitumor Chemodynamic Therapy.

We have developed a new type of nanoparticles with potent antitumor activity photoactivatable via the combination of molecular photoswitching of spiropyran (SP) and enzymatic reaction of glucose oxidase (GOx). As two key processes involved therein, Fe(III)-to-Fe(II) photoreduction in Fe(III) metal-organic frameworks (MOFs) brings about the release of free Fe2+/Fe3+ while the photoswitching of SP to merocyanine (MC) unlocks the enzymatic activity of GOx that was pre-passivated by SP. The release of free Fe3+ boosts its hydrolysis and therefore enables the acidification of microenvironment, which is further reinforced by one of the products of the GOx-mediated glucose oxidation reaction, gluconic acid (GlcA). Based on the generation of Fe2+ and acidic milieu together with another product of the oxidation reaction, hydrogen peroxide (H2O2), these two processes jointly present triple enabling factors for generating lethal hydroxyl radicals (⋅OH) species via Fenton reactions and therefore oxidative stress capable of inhibiting tumor. The antitumor potency of such nanoparticle is verified in tumor-bearing model mice in vivo, proclaiming its potential as a potent and safe agent based on the unique mechanism of optically manipulating enzyme activity for synergistic antitumor therapeutics with high spatial precision, enhanced efficacy and minimized side effects.