Tumor Research Articles

Oncometabolites in pancreatic cancer: Strategies and its implications

Pancreatic cancer (PanCa) is a catastrophic disease, being third lethal in both the genders around the globe. The possible reasons are extreme disease invasiveness, highly fibrotic and desmoplastic stroma, dearth of confirmatory diagnostic approaches and resistance to chemotherapeutics. This inimitable tumor microenvironment ( TME) or desmoplasia with excessive extracellular matrix accumulation, create an extremely hypovascular, hypoxic and nutrient-deficient zone inside the tumor. To survive, grow and proliferate in such tough TME, pancreatic tumor and stromal cells transform their metabolism. Transformed glucose, glutamine, fat, nucleotide metabolism and inter-metabolite communication between tumor and TME in synergism, impart therapy resistance, and immunosuppression in PanCa. Thus, a finer knowledge of altered metabolism would uncover its metabolic susceptibilities. These unique metabolic targets may help to device novel diagnostic/prognostic markers and therapeutic strategies for better management of PanCa. In this review, we sum up reshaped metabolic pathways in PanCa to formulate detection and remedial strategies of this devastating disease.

Exploring tumor microenvironment interactions and apoptosis pathways in NSCLC through spatial transcriptomics and machine learning.

The most common type of lung cancer is non-small cell lung cancer (NSCLC), accounting for 85% of all cases. Programmed cell death (PCD), an important regulatory mechanism for cell survival and homeostasis, has become increasingly prominent in cancer research in recent years. As such, exploring the role of PCD in NSCLC may help uncover new mechanisms for therapeutic targets. We utilized the GEO database and TCGA NSCLC gene data to screen for co-expressed genes. To delve deeper, single-cell sequencing combined with spatial transcriptomics was employed to study the intrinsic mechanisms of programmed cell death in cells and their interaction with the tumor microenvironment. Furthermore, Mendelian randomization was applied to screen for causally related genes. Prognostic models were constructed using various machine learning algorithms, and multi-cohort multi-omics analyses were conducted to screen for genes. In vitro experiments were then carried out to reveal the biological functions of the genes and their relationship with apoptosis. Cells with high programmed cell death activity primarily activate pathways related to apoptosis, cell migration, and hypoxia, while also exhibiting strong interactions with smooth muscle cells in the tumor microenvironment. Based on a set of programmed cell death genes, the prognostic model NSCLCPCD demonstrates strong predictive capabilities. Moreover, laboratory experiments confirm that SLC7A5 promotes the proliferation of NSCLC cells, and the knockout of SLC7A5 significantly increases tumor cell apoptosis. Our data indicate that programmed cell death is predominantly associated with pathways related to apoptosis, tumor metastasis, and hypoxia. Additionally, it suggests that SLC7A5 is a significant risk indicator for the prognosis of non-small cell lung cancer (NSCLC) and may serve as an effective target for enhancing apoptosis in NSCLC tumor cells.

CRISPR/RNA Aptamer System Activated by an AND Logic Gate for Biomarker-Driven Theranostics.

The development of an engineered RNA device capable of detecting multiple biomarkers to evaluate pathological states and autonomously implement responsive therapies is urgently needed. Here, we report InCasApt, an integrated nano CRISPR Cas13a/RNA aptamer theranostic platform capable of achieving both biomarker detection and biomarker-driven therapy. Within this system, a Cas13a/crRNA complex, a hairpin reporter (HR), a dinitroaniline caged Ce6 photosensitizer (Ce6-DN), and a DN-binding RNA aptamer precursor (DNBApt) are coloaded onto dendritic mesoporous silicon nanoparticles (DMSN) in a controlled manner. While InCasApt remains inert in normal cells, its programmable theranostic capabilities are activated in tumor cells that have elevated expression of carcinogenic miRNA-155 and miRNA-21. These miRNAs act as an AND logic gate, generating fluorescence for disease condition evaluation and ROS for photodynamic therapy. This process also upregulates antioncogene BRG1 and suppresses tumor migration by inhibiting the function of miRNA-155 and miRNA-21. These effects underscore the versatility of InCasApt as an miRNA-targeting strategy for bridging the gap between diagnosis and therapy.

Ultrasound nanodroplets loaded with Siglec-G siRNA and Fe3O4 activate macrophages and enhance phagocytosis for immunotherapy of triple-negative breast cancer

BackgroundThe progression of triple-negative breast cancer is shaped by both tumor cells and the surrounding tumor microenvironment (TME). Within the TME, tumor-associated macrophages (TAMs) represent a significant cell population and have emerged as a primary target for cancer therapy. As antigen-presenting cells within the innate immune system, macrophages are pivotal in tumor immunotherapy through their phagocytic functions. Due to the highly dynamic and heterogeneous nature of TAMs, re-polarizing them to the anti-tumor M1 phenotype can amplify anti-tumor effects and help mitigate the immunosuppressive TME.ResultsIn this study, we designed and constructed an ultrasound-responsive targeted nanodrug delivery system to deliver Siglec-G siRNA and Fe3O4, with perfluorohexane (PFH) at the core and mannose modified on the surface (referred to as MPFS@NDs). Siglec-G siRNA blocks the CD24/Siglec-G mediated “don’t eat me” phagocytosis inhibition pathway, activating macrophages, enhancing their phagocytic function, and improving antigen presentation, subsequently triggering anti-tumor immune responses. Fe3O4 repolarizes M2-TAMs to the anti-tumor M1 phenotype. Together, these components synergistically alleviate the immunosuppressive TME, and promote T cell activation, proliferation, and recruitment to tumor tissues, effectively inhibiting the growth of primary tumors and lung metastasis.ConclusionThis work suggests that activating macrophages and enhancing phagocytosis to remodel the TME could be an effective strategy for macrophage-based triple-negative breast cancer immunotherapy.

Metabolizable alloy clusters assemble nanoinhibitor for enhanced radiotherapy of tumor by hypoxia alleviation and intracellular PD-L1 restraint

BackgroundCancer radiotherapy (RT) still has limited clinical success because of the obstacles including radioresistance of hypoxic tumors, high-dose X-ray–induced damage to adjacent healthy tissue, and DNA-damage repair by intracellular PD-L1 in tumor.ResultsTherefore, to overcome these obstacles multifunctional core–shell BMS@Pt2Au4 nanoparticles (NPs) are prepared using nanoprecipitation followed by electrostatic assembly. Pt2Au4 clusters are released from BMS@Pt2Au4 NPs to alleviate tumor hypoxia by catalyzing the decomposition of endogenous H2O2 to generate O2 as well as by enhancing X-ray deposition at the tumor site, which thereby reduce the required X-ray dose. The released BMS-202 molecules simultaneously blockade PD-L1 on and in tumor cells, causing the activation of effector T cells and the inhibition of DNA-damage repair. Consequently, radiotherapy based on BMS@Pt2Au4 NPs enhance the expression of calreticulin on cancer cells, transposition of HMGB1 from the nucleus to the cytoplasm, generation of reactive oxygen species (ROS), DNA breakage and apoptosis of cancer cells in vitro. The tumor inhibition rate reached 92.5% under three cycles of 1-Gy X-ray irradiation in vivo.ConclusionIn conclusion, the therapeutic outcome supports the high-efficiency of radiotherapy based on BMS@Pt2Au4 NPs in hypoxic tumors expressing PD-L1.Graphical

Comparison of Shared Class I HLA-bound Non-canonical Neoepitopes between Normal and Neoplastic Tissues of Pancreatic Adenocarcinoma.

Developing T cell or vaccine therapies for pancreatic ductal adenocarcinoma (PDAC) has been challenging due to a lack of knowledge regarding immunodominant, cancer-specific antigens, as a scarcity of genomic mutation-associated neoepitopes characterizes PDAC and there are limited availability of effective approaches to discover them. We utilized an advanced mass spectrometry approach to compare the immunopeptidome of PDAC tissues and matched normal tissues from the same patients. We identified HLA class I-binding variant peptides derived from canonical proteins, which had single amino-acid substitutions not attributed to genetic mutations or RNA editing. These amino-acid substitutions appeared to result from translational errors. The variant peptides were predominantly enriched in tumor tissues, with some peptides shared across patients. Importantly, several of these variant peptides were more immunogenic than their wild-type counterparts. The shared non-canonical neoepitopes identified in this study offer promising candidates for vaccine and T cell therapy development, potentially providing new avenues for immunotherapy in PDAC.

Single Cell RNA Sequencing Before and After Light Chain Escape Reveals Intra-patient Multiple Myeloma Subpopulations with Divergent Osteolytic Gene Expression.

High-risk multiple myeloma (MM) is genomically unstable, comprised of heterogeneous populations of tumor cells that evolve over time. Light chain escape (LCE) is a clinical phenomenon observed when light chains rise separately from M-spike values, implying divergent tumor evolution. We sought to understand LCE by performing high depth transcriptomic and phenotypic studies. We performed single cell RNA sequencing and ex vivo drug sensitivity profiling on serial bone marrow biopsies from a patient with LCE at diagnosis, first relapse and relapsed/refractory timepoints. Single cell RNA sequencing uncovered distinct transcriptomic subpopulations with phenotypes that could be tracked separately by clinical serum light chain and M-spike values. Genes differentially expressed between subpopulations were assessed for generalizable effects on prognosis from the MMRF CoMMpass and GSE24080 datasets. Notably, the LCE subpopulation exhibited gene expression profile featuring prominent LAMP5-overexpression, which was associated with risk for osteolytic bone lesions. Ex vivo drug sensitivity testing displayed differential sensitivity of the subpopulations. Copy Number Variant inference showed that the transcriptomic subpopulation underlying LCE was related to a genetic subclone that evolved over time. Our findings illustrate that malignant subpopulations underly light chain escape in multiple myeloma. These studies implied that LCE and LAMP5 gene overexpression portends for increased risk of osteolytic bone disease and adverse prognosis, findings that were confirmed in the subset of patients in the CoMMpass database with light chain escape.

The combination of microwave hyperthermia with TIPE2 impedes the growth of orthotopic colon cancer.

Colon cancer (CC) is the main fatal disease of humans. Microwave hyperthermia (MH) is an adjuvant therapy for diverse cancers. Tumor necrosis factor-α induced protein-8-like 2 (TIPE2) is a tumor suppressor. However, the effect of MH combined with TIPE2 on CC remains unclear. The orthotopic CC mouse model was constructed by mouse CC CT26-Luc cells, and mice were randomized into control, model (CT26-Luc), CT26-Luc + Vector, CT26-Luc + TIPE2, CT26-Luc + MH, and CT26-Luc + MH+TIPE2 groups (n = 6). Tumor growth pretreatment and post-treatment by in vivo fluorescence image analysis was detected. TIPE2 expression and cell transfection efficiency was detected by qRT-PCR and western blot. The pathological changes by HE staining, apoptosis by TUNEL staining, serum inflammatory factors by ELISA, TIPE2 expression by immunohistochemistry, and NF-κB signaling by western blot was performed. Paracancerous tissues showed higher TIPE2 expression than in CC tissues. CT26-Luc + TIPE2, CT26-Luc + MH, and CT26-Luc + MH+TIPE2 groups reduced tumor growth, tumor cell infiltration, and increased apoptosis. CT26-Luc + TIPE2 group had lower NF-κB, TNF-α, IL-1β, IL-6, p-p65, and p-IKK expression, and elevated TIPE2 and IkB expression, which was reversed by CT26-Luc + MH group. Moreover, CT26-Luc+MH+TIPE2 group showed opposite effects on the above factor expression of CT26-Luc+MH group. Combination of MH with TIPE2 could impede CC tumor growth, providing scientific bases for its clinical application in CC treatment.

Current Combinatorial Therapeutic Aspects: The Future Prospect for Glioblastoma Treatment.

There are several types of brain tumors but glioblastoma (GBM) is one of the highly malignant tumors. A primary concern with GBM is that the treatment is inadequate. Even after giving many multi-stacked combinations of therapies to patients, inclusive of chemotherapy, radiation, and surgery, the median survival rate remains poor. Due to its heterogeneous nature, the use of selective therapy for specific targeting of tumor cells is of particular importance. Although many treatment alternatives which include surgery with adjuvant chemotherapy and radiotherapy are available, the prognosis of the disease is very poor. Combination therapy is becoming the foundation of modern antitumor therapy and it is continuously evolving and developing innovative drug regimens as evidenced by ongoing preclinical and clinical trials. In this review, we discuss the current treatment options and emerging therapeutic approaches for the treatment of GBM. The prospects for alternative glioblastoma therapy are also discussed.

Stromal immune cells expression of Siglec-15 is associated with lower T stage and better prognosis of urinary bladder cancer

IntroductionSialic acid-binding immunoglobulin-like lectin 15 (Siglec-15) is a novel immune checkpoint, similar to programmed death-ligand (PD-L1), and has emerged as a potential target for cancer immunotherapy. Until recently, little was known about the expression and role of Siglec-15 in bladder cancer (BC).MethodsIn this study, we used immunohistochemical staining to assess the expression of Siglec-15 and PD-L1 in 69 primary BC samples and analyzed their relationship with clinicopathologic characters and prognosis.ResultsThe expression rates of Siglec-15 in the tumor cells, stromal immune cells, and both the tumor and stromal cells were 84.1% (58/69), 50.7% (35/69), and 44.9% (31/69), respectively. The PD-L1 expression rate was 52.2% (36/69), with a positive rate of 17.4% (12/69). PD-L1 expression was inversely correlated with Siglec-15 expression, but the statistical significance was not achieved (P = 0.072). Low stromal Siglec-15 expression was associated with advanced tumor stage (P = 0.010). PD-L1 expression was associated with tumor stage (P = 0.008) and perineural invasion (PNI) (P = 0.048). Kaplan-Meier survival curves showed that stromal Siglec-15 expression was associated with a better prognosis (P = 0.012), although it was not an independent prognostic factor after multivariate analysis (P = 0.236) .DiscussionThis study revealed a high expression rate of Siglec-15 in BC and may provide valuable insights for patient selection in future clinical trials.

Tubulin-Based Microtentacles Aid in Heterotypic Clustering of Neutrophil-Differentiated HL-60 Cells and Breast Tumor Cells.

Circulating tumor cells (CTCs) travel through the vasculature to seed secondary sites and serve as direct precursors of metastatic outgrowth for many solid tumors. Heterotypic cell clusters form between CTCs and white blood cells (WBCs) and recent studies report that a majority of these WBCs are neutrophils in patient and mouse models. The lab discovered that CTCs produce tubulin-based protrusions, microtentacles (McTNs), which promote reattachment, retention in distant sites during metastasis and formation of tumor cell clusters. Neutrophil-CTC clusters help CTCs survive the harsh vascular environment to promote successful metastasis, however, the specific mechanism of this interaction is not fully understood. Utilizing TetherChip technology, it is found that primary and differentiated neutrophils produce McTNs composed of detyrosinated and acetylated α-tubulin and vimentin. Neutrophil McTNs aid in cluster formation, migration, and reattachment, which are suppressed with the tubulin-depolymerizing agent, Vinorelbine. Co-culturing differentiated neutrophils and tumor cells formed heterotypic clusters that enhanced migration. CTC-neutrophil clusters have higher metastatic efficiency, and by demonstrating that neutrophils form McTNs, a new possible mechanism for how neutrophils interact with tumor cells is revealed. These findings further support the idea that developing cluster-disrupting therapies can provide a new targeted strategy to reduce the metastatic potential of cancer cells.

Immunotherapy: The Next Frontier in Cancer Treatment

INTRODUCTIONOver the past decade, immunotherapy has redefined the landscape for cancer treatment, providing unprecedented survival benefits across a broad swath of tumors. The ability to harness and modulate the immune system has transformed outcomes for patients, from immune checkpoint inhibitors (ICIs) to advanced cellular therapies, such as chimeric antigen receptor (CAR) T cells and CAR macrophages (CAR-MΦ). However, these advancements have come with new challenges, such as variability in efficacy, toxicities, and lack of efficacy against the immunosuppressive tumor microenvironment (TME), especially in solid tumors[1]. In this editorial, we explore the major advances in immunotherapy, the potential of combination therapies with CAR-MΦ, and the need for these combination approaches to overcome evolving challenges. Immune Checkpoint Inhibitors: In cancers such as melanoma, NSCLC, and RCC, ICIs have become the cornerstone of immunotherapy. ICIs block inhibitory receptors such as PD-1, PD-L1, and CTLA-4, and thereby enable the immune system to overcome the suppression and unleash the immune T cells for effective anti-tumor response. In many patients, these therapies have provided durable responses and some patients have survived more than five years. We have landmark trials that show significant improvements in overall survival (OS) and progression-free survival (PFS) vs. chemotherapy in metastatic and refractory cancers[2]. Nevertheless, despite all these advances, not all patients respond to ICIs. Resistance is due to tumor antigen heterogeneity, immune evasion mechanisms, and the immunosuppressive TME. In addition, immune-mediated adverse events (images), including gastrointestinal, dermatologic, and endocrine toxicities, continue to be significant barriers. Biomarker discovery including PD-L1 expression and tumor mutational burden will become increasingly important as the field evolves for identifying patients most likely to benefit from ICIs for personalized therapy with minimal risk and cost[3]. Cellular Therapies: CAR-T Cells and the Emerging Role of CAR-MacrophagesHowever, ICIs are transformative; cellular therapies are now the new frontier in immunotherapy. Remarkable success with CAR-T cells, which involve engineering T cells to express tumor-specific receptors, has been shown in hematologic malignancies, particularly leukemia and lymphoma. Despite these barriers, however, the efficacy of these approaches in solid tumors is still limited[4]. New CAR-MΦ therapies emerging as a novel solution to meet these challenges. Chimeric antigen receptors engineered into macrophages are capable of targeting tumor cells while modifying the hostile TME. After CAR-MΦ binds to tumor cells through phagocytosis, they actively engulf tumor cells while secreting pro-inflammatory cytokines to reprogram the TME to be immunostimulatory. In addition to stimulating other immune cells like T cells and natural killer (NK) cells, these engineered macrophages also amplify anti-tumor responses[5]. Promising safety and efficacy have been demonstrated by early clinical trials, including those in HER2-expressing solid tumors. CAR-MΦ therapies can persist within the tumor, can overcome physical barriers, and can synergize with other immunotherapies. Safety issues, however, remain, most notably the possibility of cytokine release syndrome (CRS) and macrophage activation syndrome (MAS)[6]. Macrophages' intrinsic role in inflammation regulation, however, may provide a more controlled cytokine response than CAR T cells. These risks are being mitigated by tailored engineering strategies, such as IL-10 expression, to ensure safe clinical application[7]. Combination Therapies: The future of immunotherapy will be in combination with strategies to overcome its limitations and improve efficacy. In preclinical models, CAR-MΦ has synergistic activity with immune checkpoint inhibitors like anti-PD-L1 and anti-CTLA-4. Checkpoint blockade reinvigorates exhausted T cells, and CAR-MΦ remodels the TME, making it hospitable to sustained immune attack[8]. Efforts to overcome the physical and biochemical barriers within the TME are equally required. Since engineering CAR cells to secrete pro-inflammatory cytokines or enzymes that digest the extracellular matrix enhances immune cell infiltration and persistence in tumors, we hypothesized that CAR cells could be engineered to secrete diphtheria toxin, which elicits an immune response within the tumor site. Moreover, immunotherapies alone, when combined with conventional therapies like chemotherapy and radiotherapy, may increase antigen presentation, increase T cell infiltration, and increase overall immune response[9]. CONCLUSION Immunotherapy has fundamentally changed how cancer is treated and has given many patients who were once untreatable hope. However, immunosuppressive TME and durable response in solid tumors remain challenges. CAR-MΦ therapy is a promising innovation with unique advantages because they are capable of phagocytosing tumor cells, presenting antigens, and reprogramming the TME.A rational combination approach with ICIs, cellular therapies, and conventional treatments would be the future. Additionally, safety concerns need to be addressed through rigorous clinical trials and long-term follow-up to optimize treatment strategies. As we stand on the brink of this new frontier, the challenge for the scientific community is clear: Immunotherapy needs to be refined to expand its reach, to ensure it becomes a mainstay of cancer care, and to offer cures where none existed before.

NIR-715 photodynamic therapy induces immunogenic cancer cell death by enhancing the endoplasmic reticulum stress response

Effectively interfering with endoplasmic reticulum (ER) function in tumor cells and simultaneously activating an anti-tumor immune microenvironment to attack the tumor cells are promising strategies for cancer treatment. However, precise ER-stress induction is still a huge challenge. In this study, we synthesized a near-infrared (NIR) probe, NIR-715, which induces tumor cell death and inhibits tumor growth without causing apparent side effects. NIR-715 triggers severe ER stress and immunogenic cell death (ICD) after visible light exposure. NIR-715 induced ICD-associated HMGB1 release in vitro and anti-tumor immune responses, including increased cytotoxic T lymphocyte (GZMB+ CD8+ T cell) infiltration and decreased numbers of exhausted T lymphocytes (PD-L1+ CD8+ T cell). These findings suggest that NIR-715 may be a novel agent for “cold” tumor photodynamic therapy (PDT).Schematic illustration of NIR-715 photodynamic therapy for visible light-triggered, endoplasmic reticulum-targeting antitumor therapy.

Platinum–Iron Nanoparticles for Oxygen-Enhanced Sonodynamic Tumor Cell Suppression

A type of nanoparticle has been developed to simultaneously alleviate tumor hypoxia and enhance the effectiveness of sonodynamic therapy aimed at improving cancer treatment outcomes. Small-sized iron–platinum nanoparticles were prepared using a thermal reduction method, and their particle size and crystal structure were characterized. The ability of these nanoparticles to decompose hydrogen peroxide to produce oxygen and generate singlet oxygen under ultrasound irradiation was further tested. The effect of iron–platinum nanoparticles on inhibition of the proliferation of MCF-7 tumor cells under hypoxic conditions was also evaluated. The prepared iron–platinum nanoparticles effectively decomposed hydrogen peroxide to produce oxygen, reversing the hypoxic environment of tumors. Additionally, they generated singlet oxygen under ultrasound irradiation, which killed tumor cells and inhibited their proliferation. This study successfully developed small-sized iron–platinum nanoparticles that can alleviate tumor hypoxia by decomposing excess hydrogen peroxide in tumor cells to produce oxygen. Under ultrasound irradiation, these nanoparticles generate singlet oxygen, inhibiting tumor growth. The nanoparticles demonstrated good safety and are potentially valuable in enhancing oxygen-enhanced sonodynamic cancer therapy.

Engineering a controllable and reversible switch for CAR-based cellular immunotherapies via a genetic code expansion system

BackgroundAs one of the most promising adoptive cell therapies, CAR-T cell therapy has achieved notable clinical effects in patients with hematological tumors. However, several treatment-related obstacles remain in CAR-T therapy, such as cytokine release syndrome, neurotoxicity, and high-frequency recurrence, which severely limit the long-term effects and can potentially be fatal. Therefore, strategies to increase the controllability and safety of CAR-T therapy are urgently needed.MethodsIn this study, we engineered a genetic code expansion-based therapeutic system to achieve rapid CAR protein expression and regulation in response to cognate unnatural amino acids at the translational level. When the unnatural amino acid N-ε-((tert-butoxy) carbonyl)-l-lysine (BOCK) is absent, the CAR protein cannot be completely translated, and CAR-T is “closed”. When BOCK is present, complete translation of the CAR protein is induced, and CAR-T is “open”. Therefore, we investigated whether the BOCK-induced device can control CAR protein expression and regulate CAR-T cell function using a series of in vitro and in vivo experiments.ResultsFirst, we verified that the BOCK-induced genetic code expansion system enables the regulation of protein expression as a controllable switch. We subsequently demonstrated that when the system was combined with CAR-T cells, BOCK could effectively and precisely control CAR protein expression and induce CAR signaling activation. When incubated with tumor cells, BOCK regulated CAR-T cells cytotoxicity in a dose-dependent manner. Our results revealed that the presence of BOCK enables the activation of CAR-T cells with strong anti-tumor cytotoxicity in a NOG mouse model. Furthermore, we verified that the BOCK-induced CAR device provided NK cells with controllable anti-tumor activity, which confirmed the universality of this device.ConclusionsOur study systematically demonstrated that the BOCK-induced genetic code expansion system effectively and precisely regulates CAR protein expression and controls CAR-T cell anti-tumor effects in vitro and in vivo. We conclude that this controllable and reversible switch has the potential for more effective, secure, and clinically available CAR-based cellular immunotherapies.

Identification of a novel subtype of SPP1 + macrophages expressing SIRPα: implications for tumor immune evasion and treatment response prediction

BackgroundSPP1 + macrophages are among the major phagocytic cells, yet promoting tumor immune evasion and predicting unfavorable prognosis, in various cancer types. Meanwhile, the predictive value of the abundance of SPP1 + macrophages in patients receiving immunotherapy remains debatable, indicating the potential existence of subtypes of SPP1 + macrophages with diverse biological functions.MethodsThe single cell RNA sequencing data of myeloid cells integrated from several cancers including esophageal squamous cell carcinoma was analyzed for characterizing the function and cellular interactions of SPP1 + macrophages expressing SIRPα. Multiplexed immunohistochemistry was used to quantify the quantity and spatial distribution of SPP1 + macrophages expressing SIRPα. Kaplan–Meier method was used for survival analysis. In vitro and in vivo studies investigating the function of SPP1 + macrophages were performed.ResultsSPP1 + macrophages possessed a high phagocytic signature and could engulf more tumor cells in vitro and in vivo. SIRPα expression could represent the phagocytic activity of SPP1 + macrophages and delineated subsets of SPP1 + macrophages with different functions. SPP1 + SIRPα + macrophages showed close spatial distance to tumor cells and positively correlated with PD1 + CD8 + T cells. A high abundance of SPP1 + SIRPα + macrophages at baseline corresponded to patients’ response to PD-1/PD-L1 inhibitors.ConclusionA novel subtype of SPP1 + macrophages expressing SIRPα was identified and their abundance predicted patients’ response to PD-1/PD-L1 inhibitors.

Epstein-Bar Virus-Positive Lymphoepithelioma-Like Intrahepatic Cholangiocarcinoma: A Case Report and Literature Review.

Intrahepatic cholangiocarcinoma is a primary liver malignancy with poor prognosis and limited treatment options. Lymphoepithelioma-like intrahepatic cholangiocarcinoma is an exceedingly rare variant of intrahepatic cholangiocarcinoma that histologically resembles nasopharyngeal carcinoma, previously known as lymphoepithelioma. It was first reported by Hsu et al in 1996 and they were able to show the presence of Epstein-Barr virus in tumor cells. Even though several risk factors for the development of intrahepatic cholangiocarcinoma have been identified, the association between Epstein-Barr virus and intrahepatic cholangiocarcinoma is relatively rare. This article aims at discussing a female patient's lymphoepithelioma-like intrahepatic cholangiocarcinoma tumor and summarizing the existing literature on Epstein-Barr virus-positiveLymphoepithelioma-like intrahepatic cholangiocarcinoma, including the clinical features, pathogenesis, diagnostic challenges, treatment approaches, prognosis, and histopathologic description.

A spheroid whole mount drug testing pipeline with machine-learning based image analysis identifies cell-type specific differences in drug efficacy on a single-cell level

BackgroundThe growth and drug response of tumors are influenced by their stromal composition, both in vivo and 3D-cell culture models. Cell-type inherent features as well as mutual relationships between the different cell types in a tumor might affect drug susceptibility of the tumor as a whole and/or of its cell populations. However, a lack of single-cell procedures with sufficient detail has hampered the automated observation of cell-type-specific effects in three-dimensional stroma-tumor cell co-cultures.MethodsHere, we developed a high-content pipeline ranging from the setup of novel tumor-fibroblast spheroid co-cultures over optical tissue clearing, whole mount staining, and 3D confocal microscopy to optimized 3D-image segmentation and a 3D-deep-learning model to automate the analysis of a range of cell-type-specific processes, such as cell proliferation, apoptosis, necrosis, drug susceptibility, nuclear morphology, and cell density.ResultsThis demonstrated that co-cultures of KP-4 tumor cells with CCD-1137Sk fibroblasts exhibited a growth advantage compared to tumor cell mono-cultures, resulting in higher cell counts following cytostatic treatments with paclitaxel and doxorubicin. However, cell-type-specific single-cell analysis revealed that this apparent benefit of co-cultures was due to a higher resilience of fibroblasts against the drugs and did not indicate a higher drug resistance of the KP-4 cancer cells during co-culture. Conversely, cancer cells were partially even more susceptible in the presence of fibroblasts than in mono-cultures.ConclusionIn summary, this underlines that a novel cell-type-specific single-cell analysis method can reveal critical insights regarding the mechanism of action of drug substances in three-dimensional cell culture models.

Is Silver a Precious Metal for G-Quadruplex Stabilization Mediated by Porphyrins?

Cancer is a leading cause of death, so continuous efforts into cancer therapy are imperative. In tumor cells, telomerase and oncogene activity are key points for uncontrolled cell growth. Targeting these processes with ligands that inhibit telomerase and/or reduce oncogene expression has been identified as a promising cancer therapy. This study evaluated the selectivity and affinity of the silverII complex of 5,10,15,20-tetrakis(N-methyl-4-pyridinium)porphyrin (AgTMPyP) to stabilize DNA sequences capable of forming G4 structures mimicking the telomeric and oncogene regions, using spectroscopic, biochemical methods and in vitro assays. The tetracationic silver complex was compared with the free base, H2TMPyP, and the zincII complex, ZnTMPyP. The results obtained from UV-Vis and fluorescence methods pointed to a great affinity and good selectivity of AgTMPyP to G4 structures, especially for the oncogene MYC. In general, an increase in the ability of the studied ligands for 1O2 generation when interacting with oncogenic and telomeric G4 sequences was found. The results of the PCR stop assays proved that AgTMPyP has the ability to inhibit Taq polymerase. Additionally, in vitro assays demonstrated that the silverII complex exhibits low cytotoxicity against HaCaT— an immortalized, non-tumorigenic, skin keratinocytes cell line—and, although nonexclusive, AgTMPyP shows nuclear co-localization.

Sensory innervation in the prostate and a role for calcitonin gene-related peptide in prostatic epithelial proliferation

IntroductionThe prostate is densely innervated like many visceral organs and glands. However, studies to date have focused on sympathetic and parasympathetic nerves and little attention has been given to the presence or function of sensory nerves in the prostate. Recent studies have highlighted a role for sensory nerves beyond perception of noxious stimuli, as anterograde release of neuropeptides from sensory nerves can affect vascular tone and local immune responses.MethodsTo identify the degree of sensory innervation in the prostate, we utilized state-of-the-art tissue clearing and microscopy to visualize sensory innervation in the different lobes of the mouse prostate. To determine whether sensory nerves have a role in regulating proliferation within the prostate, we used an intersectional genetic and toxin approach to ablate peptidergic sensory nerves systemically.ResultsWe found that sensory neurons are abundant in the prostate both in nerve bundles along the vasculature and as independent nerve fibers wrapped around prostatic acini in a net-like fashion. In addition to the dense innervation of the prostate, we found that Calca haploinsufficiency, the genotype control for our intersectional ablation model, results in a diminished level of Ki67 staining in the stromal compartment of the dorsal lobe and a diminishing Ki67 trend in other lobes.DiscussionThese findings suggest that sensory neurons might have developmental or homeostatic effects within the prostate. Further studies are warranted to assess the role of sensory neurons and the sensory neuropeptides on prostatic development and on proliferation in the presence of pro-inflammatory stimuli such as bacterial infection or tumor cells.