Tumor Metastasis Research Articles

Onco-Ontogeny of Squamous Cell Cancer of the First Pharyngeal Arch Derivatives

Head and neck squamous cell carcinoma (H&NSCC) is an anatomic, biological, and genetic complex disease. It involves more than 1000 genes implied in its oncogenesis; for this review, we limit our search and description to the genes implied in the onco-ontogeny of the derivates from the first pharyngeal arch during embryo development. They can be grouped as transcription factors and signaling molecules (that act as growth factors that bind to receptors). Finally, we propose the term embryo-oncogenesis to refer to the activation, reactivation, and use of the genes involved in the embryo’s development during the oncogenesis or malignant tumor invasion and metastasis events as part of an onco-ontogenic inverse process.

A comprehensive review on the potential of coumarin and related derivatives as multi-target therapeutic agents in the management of gynecological cancers

Current treatments for gynecological cancers include surgery, radiotherapy, and chemotherapy. However, these treatments often have significant side effects. Phytochemicals, natural compounds derived from plants, offer promising anticancer properties. Coumarins, a class of benzopyrone compounds found in various plants like tonka beans, exhibit notable antitumor effects. These compounds induce cell apoptosis, target PI3K/Akt/mTOR signaling pathways, inhibit carbonic anhydrase, and disrupt microtubules. Additionally, they inhibit tumor multidrug resistance and angiogenesis and regulate reactive oxygen species. Specific coumarin derivatives, such as auraptene, praeruptorin, osthole, and scopoletin, show anti-invasive, anti-migratory, and antiproliferative activities by arresting the cell cycle and inducing apoptosis. They also inhibit metalloproteinases-2 and -9, reducing tumor cell migration, invasion, and metastasis. These compounds can sensitize tumor cells to radiotherapy and chemotherapy. Synthetic coumarin derivatives also demonstrate potent antitumor and anticancer activities with minimal side effects. Given their diverse mechanisms of action and minimal side effects, coumarin-class phytochemicals hold significant potential as therapeutic agents in gynecological cancers, potentially improving treatment outcomes and reducing side effects. This review will aid in the synthesis and development of novel coumarin-based drugs for these cancers.

Abstract B038: Tumor Derived Parathyroid Hormone Related Protein is an Orchestrator of Immunosuppression in Pancreatic Cancer

Abstract Pancreatic ductal adenocarcinoma (PDAC) has the lowest 5-year survival of common cancers, owing to a lack of early detection and ineffective therapies. Immune checkpoint inhibitors hold promise for PDAC patients, and while they represent the most ongoing clinical trials, they have had little success. Parathyroid Hormone Related Protein (PTHrP; encoded by PTHLH) defines the most aggressive squamous/basal/quasi-mesenchymal PDAC subtype. Our previous work revealed autocrine PTHrP signaling mediates tumor growth, epithelial-to-mesenchymal transition (EMT), and metastasis. However, PTHrP’s relationship with immune cells of the tumor microenvironment (TME) remains unknown. Investigation of tumors from KrasG12D; p53R172H; Pdx1-Cre; YFP (KPCY) and KPCY-Pthlh loxP/WT (KPCY-Pthlhhet) mice revealed less tumor associated neutrophils (TANs) and greater cytotoxic T cell infiltration, demonstrating the necessity of PTHrP for immune exclusion. Orthotopic implantation of Pthlh overexpressing KPCY cells (KPCY-PthlhOE) revealed a greater proportion of TANs than the control. A concomitant decrease in the number of effector T and natural killer cells in the KPCY-PthlhOE tumors was observed, showing PTHrP is sufficient to organize an immunosuppressive TME. Interestingly, recombinant human PTHrP was sufficient to induce TANs mobilization in vivo. To interrogate the mechanism of neutrophil trafficking into the TME, cytokine production was quantified in vitro. KPCY-PthlhOE exhibited a pronounced increase of Cxcl1 expression, a strong neutrophil chemokine, leading us to believe neutrophils migrate to the TME using a Cxcl1 gradient. Therefore, we hypothesized that PTHrP-mobilized neutrophils express the receptor for CXCL1; CXCR2. Indeed, recombinant PTHrP was sufficient to mobilize Cxcr2+ neutrophils in vivo. Furthermore, recombinant PTHrP was able to induce Cxcl1 production by KPCY tumor cells in vitro in a cAMP response element-binding protein(CREB)-dependent manner. These data describe a novel mechanism for the mobilization and infiltration of immune suppressive TANs into the TME, initiated by PTHrP. The PTHrP-dependent immunosuppressive TME excludes and inhibits effector immune cells. Further interrogation of PTHrP signaling during the genesis of the immune suppressive TME in PDAC may lead to the development of therapeutics to meet this unmet clinical need. Citation Format: Calvin Johnson, Jason Pitarresi. Tumor Derived Parathyroid Hormone Related Protein is an Orchestrator of Immunosuppression in Pancreatic Cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B038.

Abstract C074: Influence of Cell Death and Stress on the Tumor Microenvironment and EMT in Pancreatic Ductal Adenocarcinoma Through Single Cell and Spatial Transcriptomics

Abstract Pancreatic cancer remains one of the most deadly cancers, with over 450,000 fatalities in 2022 alone. The most lethal variation of this cancer is Pancreatic Ductal Adenocarcinoma (PDAC), with a 5-year survival rate of under 10%. Combination chemotherapy has provided major advances in this disease but it remains challenging for many reasons including the immunosuppressive nature of the tumor microenvironment (TME). PDAC has a highly desmoplastic microenvironment that suppresses antitumor immunity and promotes tumor cell invasion, metastasis, and resistance through epithelial to mesenchymal transition (EMT) of cancer cells. Cancer cells can undergo various forms of cell death, notably apoptosis, necrosis, autophagy, pyroptosis, and ferroptosis. These mechanisms are triggered by different cellular stressors and shape the microenvironment. Through the use of 15 Visium spatial transcriptomics samples from publicly available primary PDAC tumors and paired single-cell RNA (scRNA) data, we aim to understand how cell stress and cell death shape the tumor microenvironment (TME) and epithelial-mesenchymal transition (EMT). By leveraging information from known pathways in literature, we will identify cellular programs related to stress, death, and EMT. We will explore intracellular correlations within single-cell data and extracellular interactions within the TME using spatial data to uncover how different forms of cell death and stress signatures influence the TME and EMT spectrum. The initial phase involves identifying the types of stress that lead to various forms of cell death intracellularly and validating these gene signatures. Subsequently, we will apply these validated signatures to the spatial data to correlate each score and search for significant overlapping programs. This will extend beyond stress and death, incorporating TME signatures corresponding to different cell types and the EMT spectrum. By understanding the signals that promote the death of various cell populations, we aim to identify combination treatments that target both ends of the EMT spectrum. Additionally, this research will provide insights into the inflammatory impact of cell death on the TME. Our approach aims to inform the development of therapies that disrupt resistance mechanisms, ultimately improving treatment outcomes for patients with PDAC. This research can potentially reveal new therapeutic targets and strategies, enhancing our ability to combat this deadly disease. Citation Format: Rahul Bansal, Izabella Zamora, Samuel Wright, Nir Hacohen, Arnav Mehta. Influence of Cell Death and Stress on the Tumor Microenvironment and EMT in Pancreatic Ductal Adenocarcinoma Through Single Cell and Spatial Transcriptomics [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C074.

Abstract A041: Deciphering the bidirectional influences of sympathetic neurons and cancer associated fibroblasts and the resulting contributions to PDAC progression

Abstract The pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment (TME) is innervated by a variety of peripheral nerves that are emerging as facilitators of tumor initiation, progression and metastasis. However, the influences of sympathetic nerves on the TME and how these bi-directional interactions can exacerbate PDAC progression remains unclear. Sympathetic nerves innervate both human and murine PDAC tumors and are frequently surrounded by abundant and heterogeneous cancer-associated fibroblasts (CAFs), which are known to play various crucial roles in PDAC progression. In our studies, we aim to elucidate the bidirectional CAF and sympathetic-neuron crosstalk that may promote tumor growth using a variety of in vitro coculture systems to model sympathetic-CAF interactions, as well as in vivo modeling of nerves in murine PDAC tumors. Our nerve-CAF cocultures reveal that CAFs closely and frequently interact with sympathetic nerves, and also enhance sympathetic nerve outgrowth. We further sequenced these cocultured conditions to evaluate intrinsic transcriptional changes that may be contributing to the enhanced nerve outgrowth. We identified several genes that may contribute to upregulated CAF adhesion, ECM deposition and neuroactive ligand-receptor interactions, which may also be supportive of the axon outgrowth and tumor cell growth. We are also currently characterizing human and murine PDAC samples that contain and lack sympathetic innervation to evaluate spatial phenotypic shifts within the TME. Collectively, our results suggest that this crosstalk between sympathetic neurons and CAFs may both enhance tumor innervation and tumor-promoting phenotypes in the TME, such as axonogenesis and fibrosis in PDAC. Citation Format: Ariana Sattler, Parham Diba, Tetiana Korzun, Marigold Kuykendall, Kevin MacPherson-Hawthorne, Mara Sherman, Teresa Zimmers, Sebnem E Eksi. Deciphering the bidirectional influences of sympathetic neurons and cancer associated fibroblasts and the resulting contributions to PDAC progression [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A041.

Abstract A058: Primary cancer-associated fibroblasts alter tumor organoid chemosensitivity through epigenetic regulation

Abstract Pancreatic adenocarcinoma (PDAC) is the 3rd leading cause of cancer-related deaths in Canada with a 5-year survival rate at ∼12%. A major cause of this poor survival rate is the inefficiency of current treatments. Only 20% of patients are eligible for surgical resection while the other 80% are treated with chemotherapies (nab-paclitaxel, Gemcitabine, FOLFIRINOX) having limited efficacy. This resistance is due to the PDAC tumour microenvironment (TME), including cancer-associated fibroblasts (CAFs). Numerous studies show CAFs and the TME affect tumour growth, chemoresistance and metastasis, but the mechanisms underlying the effects are unknown. Epigenetic mechanisms, including DNA methylation and histone modifications, link environmental factors to altered gene expression. Genes encoding epigenetic mediators are often mutated or over-expressed in PDAC, but a role in resistance has yet to be defined. We hypothesize that epigenetic reprogramming occurs in pancreatic cancer cells in response to secreted factors from CAFs leading to increased therapy resistance. Methods: Most work to date on PDAC resistance has been performed with immortalized cell lines and in two dimensions. Therefore, to test our hypothesis, we established patient-derived organoids (PDO) and CAFs obtained from endoscopic ultrasound (EUS). Clinical data was obtained through the DERIVE (Determination of Response to Therapy in Individual Patients) database. To examine the effects of CAFs on PDOs, we obtained conditioned media (CM) from four, early passage CAFs, grown 48 hours in culture. Organoid media was supplemented 1:1 with CAF-CM and six PDOs grown in this media for up to 14 days. PDO growth and the response to gemcitabine, FOLFIRINOX or radiation was monitored using an Incucyte system. To identify changes in the epigenome and how this reflects changes in gene expression, DNA methylation arrays and ATAC-Seq was performed before and after treatment with CAF-CM and aligned to RNA-seq analysis on RNA isolated at the same time point. Results: Exposure of PDOs to CAF-CM resulted in altered phenotypes and molecular profiles of PDOs in a patient-specific fashion. Increased resistance to chemotherapy (Gemcitabine) and radiotherapy was observed in multiple PDOs. Changes in DNA methylation and chromatin accessibility was aligned with transcriptomic data and identified known and novel pathways linked to chemoresistance. Metabolomic and cytokine analysis on CAFs-CM identified differential enrichment of metabolites linked to epigenetic mediators. We are currently determining if these factors secreted by the CAFs promote chemo and radio-resistance. Conclusions: This study shows the importance in integrating TME components when examining novel therapeutic options. In addition, we show (1) CAFs secrete factors that enhance therapeutic resistance in PDOs and (2) these changes likely involve altered epigenetic reprogramming. Future experiments will identify the epigenetic factors that can be targeted to counteract chemoresistance. Citation Format: Emilie Jaune-Pons, Zachary Klassen, Joana Ribeiro Pinto, Maria Nica, Nadeem Hussain, Michael Sey, Ken Leslie, Matthew Cecchini, Crystal Engelage, Danielle Porplycia, Stephen Welch, Brian Yan, Christopher Pin. Primary cancer-associated fibroblasts alter tumor organoid chemosensitivity through epigenetic regulation [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A058.

Abstract C005: Exploratory biomarker evaluation of the randomized Phase 2 cohort of CM24 in combination with nivolumab and chemotherapy in advanced/metastatic pancreatic cancer

Abstract Background: CM24 (mAb) blocks CEACAM1 interactions, has key roles in cancer progression, immune escape and metastasis. CEACAM1 is part of the neutrophil extracellular trap (NET) complex, involved in tumor immune evasion, metastasis and cancer-associated thrombosis, affecting patient survival. We demonstrated that CM24 binds to NET and suppresses NET-induced migration of cancer cells (AACR Cancer Metastasis Nov.2022) and that CM24+nivolumab (nivo) treatment of pancreatic ductal adenocarcinoma (PDAC) patients markedly reduced serum levels of the NET marker, Myeloperoxidase (MPO; AACR on pancreatic cancer Sept.2023). In a Phase 2 study (NCT04731467) 31 patients with advanced/metastatic PDAC progressed after 1st line therapy, were randomized for CM24 (20mg/kg q2wk), nivo (240mg/kg q2wk) and liposomal irinotecan, 5 fluorouracil and leucovorin (Nal-IRI/5FU/LV, q2wk) treatment (experimental arm) vs Nal-IRI/5FU/LV only (control arm). Interim analysis of the data showed Median OS for the experimental arm was 7.72m (95% CI: 4.00-8.11) vs. 5.62m (95% 3.22 -7.89) for the control arm (HR: 0.74; 95% CI: 0.31-1.77). Median PFS for the experimental group was 3.8m (95% 1.8-5.0) vs. 1.9m (95% 0.9, 3.6) for the control arm (HR 0.72; 95% CI: 0.33-1.60) (ASCO2024-LBA4143). Here we present a subgroup analysis evaluating serum MPO as a potential biomarker for CM24-based therapy. Methods: Patients were randomized as described above. Pre-dose serum samples were collected from 30 out of 31 pts and serum MPO levels were evaluated using ELISA (Cayman), where the measured mean serum MPO in healthy subjects (n=30) was 53 +/-7[SE]. NET-induced blood coagulation was tested using platelet aggregation assay. Results: Mean pre-dose MPO level in patients (n=30) was 359 +/- 46 [SE] and was selected as a threshold for biomarker analysis. OS for patients with MPO levels below the threshold (n=9/16) was 8.1 mo in the experimental arm (HR 0.34; (95% CI: 0.099-1.149)) vs 4.5mo in the control (n=9/15). PFS for patients with MPO levels below the threshold was 4.2mo in the experimental arm vs. 1.7 mo in the control (HR 0.24 (95% CI: 0.20-1.59)). NET-associated blood coagulation and the inhibitory effect of CM24 were tested in vitro. NETs induced platelet aggregation in a dose-dependent manner, and CM24 significantly reduced it (50-70% reduction, p value < 0.0001) at NET levels related to the selected biomarker range (MPO<357) but was less effective at higher NET levels (MPO>500), which relate to MPO above the threshold. These results are in line with the proposed CM24 MoA, affecting NET-related cancer complications. Conclusions: Exploratory biomarker analysis suggests serum MPO as a potential predictive biomarker for CM24-based therapy showing prolonged median OS, PFS and 66% reduction in risk of death vs control in PDAC patients with MPO levels below threshold compared to interim analysis that estimated OS and PFS risk reduction of 26% and 28%, respectively. These findings will be further evaluated in upcoming clinical studies. Citation Format: Teresa Macarulla, Michael Cecchini, Rocio Garcia-Carbonero, Talia Golan, Ruth Perets, Erkut Borazanci, Manuel Pedegral, Mariano Ponz-Sarvisé, Mohammed Najeeb Al Hallak, Shubham Pant, Valentina Boni, Omar Saavedra, María José de Miguel-Luken, Alexis D. Leal, Andrés J. Muñoz Martín, Tamara Sauri-Nadal, Michael Schickler, Hava Ben David, Hadas Reuveni. Exploratory biomarker evaluation of the randomized Phase 2 cohort of CM24 in combination with nivolumab and chemotherapy in advanced/metastatic pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C005.

Deacetylation by SIRT6 increases the stability of GILZ to suppress NSCLC cell migration and invasion

Glucocorticoid-induced leucine zipper (GILZ) plays a role in cancer cell proliferation in several tumor types. However, in our present study, GILZ was demonstrated to be a metastasis regulator but not a proliferation regulator in non-small cell lung cancer (NSCLC). The overexpression of GILZ had no significant effect on the proliferation of NSCLC cells but inhibited their metastasis by targeting the epithelial-mesenchymal transition pathway. The deacetylase SIRT6, a key regulator of protein stability, can enhance the stability of the GILZ protein by mediating its deacetylation, which prevents ubiquitination and degradation. This process ultimately enhances the inhibitory effect of GILZ on the migration and invasion of NSCLC cells. Thus, GILZ may be a promising new therapeutic target for tumor metastasis.

Tumor to tumor metastasis of renal cell carcinoma in carcinoma stomach: An unusual case presentation

Tumor to tumor metastasis of renal cell carcinoma in carcinoma stomach: An unusual case presentation

Clinical significance of peripheral blood UL16 and DR-70 for the early diagnosis and prognostic evaluation of colorectal cancer

BACKGROUND Early diagnosis of colorectal cancer (CRC) is of great significance to improve the survival rate and quality of life of patients, but early diagnosis of CRC requires more sensitive techniques. Peripheral blood UL16-binding protein 2 (ULBP2) and human fibrinogen degradation products (DR-70) are the main indicators for the diagnosis of malignant tumors. AIM To assess ULBP2 and DR-70 potential for the early diagnosis and prognostic evaluation of CRC to provide a reference. METHODS This study involved 60 patients with early-stage CRC (CRC group), 50 patients with benign colorectal tumors (benign group), and 50 healthy patients (control group) enrolled at the Affiliated Hospital of Jiangnan University and Jiangsu Province Official Hospital between January, 2020 and January, 2022. ULBP2 and DR-70 levels in the blood were determined and differences among the three groups and early diagnostic values for CRC were determined. Patients with CRC were divided into the good prognosis and poor prognosis groups, and ULBP2 and DR-70 levels in the blood and diagnostic values were compared. RESULTS ULBP2 and DR-70 serum levels were significantly higher in the CRC group than in the control and benign groups (P < 0.05); however, no significant differences were observed between the benign and control groups (P > 0.05). Among the 60 patients with CRC followed up for two years, two died (3.33%) and 15 exhibited tumor metastasis, progression, or recurrence (25.00%). ULBP2 and DR-70 serum levels were significantly higher in the poor prognosis group than in the good prognosis group (P < 0.05). A receiver operating characteristic curve was plotted. Area under the curve, sensitivity, and specificity of serum ULBP2 with DR-70 for the early diagnosis of CRC were higher than those of the single serum indices (P < 0.05) in both the good and poor prognosis groups. CONCLUSION ULBP2 and DR-70 serum levels were significantly high in patients with early-stage CRC. They improved the diagnostic rate of early-stage CRC and predicted patient prognosis, thereby showing clinical application potential.

Recombinant ferritin-based nanoparticles as neoantigen carriers significantly inhibit tumor growth and metastasis

BackgroundTumor neoantigen peptide-based vaccines, systemic immunotherapies that enhance antitumor immunity by activating and expanding antigen-specific T cells, have achieved remarkable results in the treatment of a variety of solid tumors. However, how to effectively deliver neoantigens to induce robust antitumor immune responses remains a major obstacle.ResultsHere, we developed a safe and effective neoantigen peptide delivery system (neoantigen-ferritin nanoparticles, neoantigen-FNs) that successfully achieved effective lymph node targeting and induced robust antitumor immune responses. The genetically engineered self-assembled particles neoantigen-FNs with a size of 12 nm were obtained by fusing a neoantigen with optimized ferritin, which rapidly drainage to and continuously accumulate in lymph nodes. The neoantigen-FNs vaccine induced a greater quantity and quality of antigen-specific CD8+ T cells and resulted in significant growth control of multiple tumors, dramatic inhibition of melanoma metastasis and regression of established tumors. In addition, no obvious toxic side effects were detected in the various models, indicating the high safety of optimized ferritin as a vaccine carrier.ConclusionsHomogeneous and safe neoantigen-FNs could be a very promising system for neoantigen peptide delivery because of their ability to efficiently drainage to lymph nodes and induce efficient antitumor immune responses.Graphical

ING5 inhibits aerobic glycolysis of lung cancer cells by promoting TIE1-mediated phosphorylation of pyruvate dehydrogenase kinase 1 at Y163.

Aerobic glycolysis is critical for tumor growth and metastasis. Previously, we have found that the overexpression of the inhibitor of growth 5 (ING5) inhibits lung cancer aggressiveness and epithelial-mesenchymal transition (EMT). However, whether ING5 regulates lung cancer metabolism reprogramming remains unknown. Here, by quantitative proteomics, we showed that ING5 differentially regulates protein phosphorylation and identified a new site (Y163) of the key glycolytic enzyme PDK1 whose phosphorylation was upregulated 13.847-fold. By clinical study, decreased p-PDK1Y163 was observed in lung cancer tissues and correlated with poor survival. p-PDK1Y163 represents the negative regulatory mechanism of PDK1 by causing PDHA1 dephosphorylation and activation, leading to switching from glycolysis to oxidative phosphorylation, with increasing oxygen consumption and decreasing lactate production. These effects could be impaired by PDK1Y163F mutation, which also impaired the inhibitory effects of ING5 on cancer cell EMT and invasiveness. Mouse xenograft models confirmed the indispensable role of p-PDK1Y163 in ING5-inhibited tumor growth and metastasis. By siRNA screening, ING5-upregulated TIE1 was identified as the upstream tyrosine protein kinase targeting PDK1Y163. TIE1 knockdown induced the dephosphorylation of PDK1Y163 and increased the migration and invasion of lung cancer cells. Collectively, ING5 overexpression-upregulated TIE1 phosphorylates PDK1Y163, which is critical for the inhibition of aerobic glycolysis and invasiveness of lung cancer cells.

Research on the biophysical properties of circulating tumor cells

This article reviews the biophysical properties of circulating tumor cells (CTCs) and their significance in tumor metastasis and liquid biopsy. CTCs are tumor cells that have shed from solid tumors or metastatic foci and entered the bloodstream, possessing various biological markers. They may be cleared by the immune system, enter a dormant state, or form clusters to facilitate metastasis. The physical properties of CTCs, including cell size, density, rigidity, and dielectric properties, differ from normal blood cells, providing a theoretical basis for separation and detection. The biological characteristics of CTCs include epithelial cell properties, expression of specific surface marker proteins, and the characteristics of epithelial-mesenchymal transition, which are crucial for the aggressiveness and metastatic potential of tumors. The development of CTC detection technologies, such as the Cell Search system, has been clinically applied, but the heterogeneity of tumor cells remains a major challenge in research. The development of composite separation technologies that combine various properties of CTCs is expected to improve the sensitivity, specificity, and efficiency of CTC detection, thereby playing an important role in the personalized treatment and prognostic assessment of tumors.

Phytochemicals and Nanotechnology: A Powerful Combination against Breast Cancer.

Considerable advancements have been made in breast cancer therapeutics in the past few decades. However, the advent of chemo-resistance and adverse drug reactions coupled with tumor metastasis and recurrence posed a serious threat to combat this lethal disease. Novel anti-cancer agents, as well as new therapeutic strategies, are needed to complement conventional breast cancer therapies. The quest for developing novel anti-cancer drugs caused an upsurge in exploring and harnessing natural compounds, especially phytochemicals. Various research groups have explored and documented the anti-cancer potential of wide variety of phytochemical groups including flavonoids (curcumin, kaempferol, myricetin, quercetin, naringenin, apigenin, genistein epigallocatechin gallate), stilbenes (resveratrol), carotenoids (crocin, lycopene, lutein), and anthraquinone (Emodin). However, low chemical stability, poor water solubility, and short systemic half-life impede their clinical utility. The implication of nano-technological approaches to decode the pharmacokinetic challenges associated with phytochemical usage, as well as selective drug targeting, have markedly enhanced the pre-clinical anti-cancer activity, thus aiding in their clinical translation. This review documented the recent advances in utilizing phytochemicals for breast cancer prevention and lipidbased nanotechnological approaches for circumventing their pharmacokinetic concerns to enhance their systemic availability, cytotoxicity, and targeted delivery against breast cancer alone as well as in combination with conventional therapeutic agents.

Towards imaging the immune state of cancer by PET: Targeting legumain with 11C-labeled P1-Asn peptidomimetics carrying a cyano-warhead

PurposeM2-type tumor-associated macrophages (TAM) residing in the tumor microenvironment (TME) have been linked to tumor invasiveness, metastasis and poor prognosis. M2 TAMs suppress T cell activation, silencing the recognition of the cancer by the immune system. Targeting TAMs in anti-cancer therapy may support the immune system and immune-checkpoint inhibitor therapies to fight the cancer cells. We aimed to develop a PET tracer for the imaging of M2 TAM infiltration of cancer, using activated legumain as the imaging target. Basic proceduresTwo P1-mimicking inhibitors with a cyano-warhead were labeled with carbon-11 and evaluated in vitro and in vivo with a CT26 tumor mouse model. Target expression and activity were quantified from RT-qPCR and in vitro substrate conversion, respectively. The co-localization of legumain and TAMs was assessed by fluorescence microscopy. The two tracers were evaluated by PET with subsequent biodistribution analysis with the dissected tissues. Parent-to-total radioactivity in plasma was determined at several time points after i.v. tracer injection, using reverse phase radio-UPLC. Main findingsLegumain displayed a target density of 40.7 ± 19.1 pmol per mg total protein in tumor lysate (n = 4) with high substrate conversion and colocalization with M2 macrophages in the tumor periphery. [11C]1 and [11C]2 were synthesized with >95 % radiochemical purity and 12.9–382.2 GBq/μmol molar activity at the end of synthesis. We observed heterogeneous tumor accumulation in in vitro autoradiography and PET for both tracers. However, excess unlabeled 1 or 2 did not compete with tracer accumulation. Both [11C]1 and [11C]2 were rapidly metabolized to a polar radiometabolite in vivo. Principal conclusionsThe legumain tracers [11C]1 and [11C]2, synthesized with high radiochemical purity and molar activity, accumulate in the legumain-positive CT26 tumor in vivo. However, the lack of competition by excess compound questions their specificity. Both tracers are rapidly metabolized in vivo, requiring structural modifications towards more stable tracers for further investigations.

Long non-coding RNA LNC-POTEM-4 promotes HCC progression via the LNC-POTEM-4/miR-149-5p/Wnt4 signaling axis

Information on the potential role of the long non-coding RNA LNC-POTEM-4 in cancer progression is limited. Our preliminary study found that LNC-POTEM-4 was overexpressed in hepatocellular carcinoma (HCC) tissues, which led us to further investigate the biological function and molecular mechanism of LNC-POTEM-4 in HCC development. LNC-POTEM-4 expression in HCC tissues was examined using transcriptome sequencing and quantitative reverse transcription PCR. The relationships between LNC-POTEM-4 and the stage and prognosis of HCC in patient data from the TCGA database were analyzed. The effects of LNC-POTEM-4 on proliferation, invasion/migration, and epithelial-mesenchymal transition marker expression in HCC cells were evaluated in vitro using gain- and loss-of-function assays, while its effects on tumor growth and metastasis were explored through animal experiments. A LNC-POTEM-4/microRNA (miR)-149-5p/Wnt4 regulatory signaling axis was identified using bioinformatics analysis, and dual luciferase reporter, RNA immunoprecipitation, and RNA pull-down assays. Co-transfection of LNC-POTEM-4 and Wnt4 expression plasmids was employed to confirm the new signaling pathway. We found that LNC-POTEM-4 was overexpressed in HCC tissues and was linked to poor staging and prognosis. LNC-POTEM-4 promoted proliferation, invasion, migration, and the epithelial-mesenchymal transition of HCC cells in vitro. Silencing of LNC-POTEM-4 inhibited HCC growth and distant metastasis in vivo. Mechanically, LNC-POTEM-4 was found to function as a competitive endogenous RNA, upregulating Wnt4 by sponging miR-149-5p to promote HCC progression. Wnt4 overexpression may have counteracted the tumor-inhibition effect of LNC-POTEM-4 silencing. In conclusion, LNC-POTEM-4 upregulated Wnt4 to activate the Wnt signaling pathway and stimulate the malignancy tendency of HCC by sponging miR-149-5p, providing a prospective target for the detection and therapy of HCC. However, the effects of LNC-POTEM-4 on the miR-149-5p/Wnt4 signaling axis should be further studied in animal experiments.

Exosomal miR-4745-5p/3911 from N2-polarized tumor-associated neutrophils promotes gastric cancer metastasis by regulating SLIT2

Tumor cells remodel the phenotype and function of tumor microenvironment (TME) cells to favor tumor progression. Previous studies have shown that neutrophils in TME are polarized to N2 tumor-associated neutrophils (TANs) by tumor derived factors, thus promoting tumor growth and metastasis, angiogenesis, therapy resistance, and immunosuppression. Exosomes act as critical intercellular messengers in human health and diseases including cancer. So far, the biological roles of exosomes from N2 TANs in gastric cancer have not been well characterized. Herein, we represented the first report that exosomes from N2 TANs promoted gastric cancer metastasis in vitro and in vivo. We found that exosomes from N2 TANs transferred miR-4745-5p/3911 to gastric cancer cells to downregulate SLIT2 (slit guidance ligand 2) gene expression. Adenovirus-mediated overexpression of SLIT2 reversed the promotion of gastric cancer metastasis by N2 TANs derived exosomes. We further revealed that gastric cancer cells induced glucose metabolic reprogramming in neutrophils through exosomal HMGB1 (high mobility group protein B1)/NF-κB pathway, which mediated neutrophil N2 polarization and miR-4745-5p/3911 upregulation. We further employed ddPCR (droplet digital PCR) to detect the expression of miR-4745-5p/3911 in N2 TANs exosomes from human serum samples and found their increased levels in gastric cancer patients compared to healthy controls and benign gastric disease patients. Conclusively, our results indicate that N2 TANs facilitate cancer metastasis via regulation of SLIT2 in gastric cancer cells by exosomal miR-4745-5p/3911, which provides a new insight into the roles of TME cells derived exosomes in gastric cancer metastasis and offers a potential biomarker for gastric cancer diagnosis.

Focal adhesion kinase-mediated interaction between tumor and immune cells in the tumor microenvironment: implications for cancer-associated therapies and tumor progression.

Focal adhesion kinase (FAK) expression has been linked to tumor growth, immunosuppression, metastasis, angiogenesis, and therapeutic resistance through kinase-dependent and kinase scaffolding functions in the nucleus and cytoplasm. Hence, targeting FAK alone or with other agents has gained attention as a potential therapeutic strategy. Moreover, mounting evidence shows that FAK activity can influence the tumor immune microenvironment crosstalk to support tumor progression. Recently, tumor immune microenvironment interaction orchestrators have shown to be promising therapeutic agents for cancer immunotherapies. Therefore, this review highlights how FAK regulates the tumor immune microenvironment interplay to promote tumor immune evasive mechanisms and their potential for combination therapies with standard cancer treatments.

Oxidative Stress and Age-Related Tumors

Oxidative stress is the result of the imbalance between reactive oxygen and nitrogen species (RONS), which are produced by several endogenous and exogenous processes, and antioxidant defenses consisting of exogenous and endogenous molecules that protect biological systems from free radical toxicity. Oxidative stress is a major factor in the aging process, contributing to the accumulation of cellular damage over time. Oxidative damage to cellular biomolecules, leads to DNA alterations, lipid peroxidation, protein oxidation, and mitochondrial dysfunction resulting in cellular senescence, immune system and tissue dysfunctions, and increased susceptibility to age-related pathologies, such as inflammatory disorders, cardiovascular and neurodegenerative diseases, diabetes, and cancer. Oxidative stress-driven DNA damage and mutations, or methylation and histone modification, which alter gene expression, are key determinants of tumor initiation, angiogenesis, metastasis, and therapy resistance. Accumulation of genetic and epigenetic damage, to which oxidative stress contributes, eventually leads to unrestrained cell proliferation, the inhibition of cell differentiation, and the evasion of cell death, providing favorable conditions for tumorigenesis. Colorectal, breast, lung, prostate, and skin cancers are the most frequent aging-associated malignancies, and oxidative stress is implicated in their pathogenesis and biological behavior. Our aim is to shed light on the molecular and cellular mechanisms that link oxidative stress, aging, and cancers, highlighting the impact of both RONS and antioxidants, provided by diet and exercise, on cellular senescence, immunity, and development of an antitumor response. The dual role of ROS as physiological regulators of cell signaling responsible for cell damage and diseases, as well as its use for anti-tumor therapeutic purposes, will also be discussed. Managing oxidative stress is crucial for promoting healthy aging and reducing the risk of age-related tumors.

FOXF1 inhibits invasion and metastasis of lung adenocarcinoma cells and enhances anti-tumor immunity via MFAP4/FAK signal axis

Based on the joint analysis of multi-omic data and the biological experiments, we demonstrate that FOXF1 inhibits invasion and metastasis of lung adenocarcinoma cells and enhances anti-tumor immunity via regulating MFAP4/FAK signal axis in this study. The levels of FOXF1 and MFAP4 are significantly down-regulated in LUAD, and the increased levels of two genes can improve the clinical prognosis of LUAD patients. Fluorescein reporter gene determination, chromatin immunoprecipitation and gene co-expression analysis indicate that MFAP4 level is positively regulated by transcription factor FOXF1. The function enrichment analysis shows that the levels of FOXF1 and MFAP4 are closely associated with an enrichment of tumor metastasis signatures. FOXF1 can inhibit the migration and invasion of LAUD cells by transcriptionally activating MFAP4 expression. And the overexpression of FOXF1/MFAP4 can reduce focal adhesion kinase (FAK) phosphorylation, while their knockdown result in the opposite effects. The increased levels of FOXF1/MFAP4 enhance the antitumor immunity by increasing the infiltration of dendritic cells and CD4+ T cells, and the interactions between LUAD cells and immune cells, and activating multiple anti-tumor immunity-related pathways. In conclusion, our study reveals the potential function of FOXF1/MFAP4/FAK signal axis in inhibiting metastasis of LUAD cells and modulating anti-tumor immunity of LUAD patients.