Mesoporous manganese nanocarrier target delivery metformin for the co-activation STING pathway to overcome immunotherapy resistance.

Targeting the stimulator of interferon genes (STING) pathway is a promising strategy to overcome primary resistance to immune checkpoint inhibitors in non-small cell lung cancer with the STK11 mutation. We previously found metformin enhances the STING pathway and thus promotes immune response. However, its low concentration in tumors limits its clinical use. Here, we constructed high-mesoporous Mn-based nanocarrier loading metformin nanoparticles (Mn-MSN@Met-M NPs) that actively target tumors and respond to release higher concentration of Mn2+ ions and metformin. The NPs significantly enhanced the T cells to kill lung cancer cells with the STK11 mutant. The mechanism shows that enhanced STING pathway activation promotes STING, TBKI, and IRF3 phosphorylation through Mn2+ ions and metformin release from NPs, thus boosting type I interferon production. In vivo, NPs in combination with a PD-1 inhibitor effectively decreased tumor growth. Collectively, we developed a Mn-MSN@Met-M nanoactivator to intensify immune activation for potential cancer immunotherapy.

Understanding the dynamics of TKI-induced changes in the tumor immune microenvironment for improved therapeutic effect.

BACKGROUND: The dynamic interplay between tyrosine kinase inhibitors (TKIs) and the tumor immune microenvironment (TME) plays a crucial role in the therapeutic trajectory of non-small cell lung cancer (NSCLC). Understanding the functional dynamics and resistance mechanisms of TKIs is essential for advancing the treatment of NSCLC. METHODS: This study assessed the effects of short-term and long-term TKI treatments on the TME in NSCLC, particularly targeting epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) mutations. We analyzed changes in immune cell composition, cytokine profiles, and key proteins involved in immune evasion, such as laminin subunit γ-2 (LAMC2). We also explored the use of aspirin as an adjunct therapy to modulate the TME and counteract TKI resistance. RESULTS: Short-term TKI treatment enhanced T cell-mediated tumor clearance, reduced immunosuppressive M2 macrophage infiltration, and downregulated LAMC2 expression. Conversely, long-term TKI treatment fostered an immunosuppressive TME, contributing to drug resistance and promoting immune escape. Differential responses were observed among various oncogenic mutations, with ALK-targeted therapies eliciting a stronger antitumor immune response compared with EGFR-targeted therapies. Notably, we found that aspirin has potential in overcoming TKI resistance by modulating the TME and enhancing T cell-mediated tumor clearance. CONCLUSIONS: These findings offer new insights into the dynamics of TKI-induced changes in the TME, improving our understanding of NSCLC challenges. The study underscores the critical role of the TME in TKI resistance and suggests that adjunct therapies, like aspirin, may provide new strategies to enhance TKI efficacy and overcome resistance.

Overall signature of acquired KRAS gene changes in advanced non-small cell lung cancer patient with EGFR-TKI resistance.

OBJECTIVE: Numerous scattered case studies continue to demonstrate a strong correlation between acquired KRAS mutations and epidermal growth factor receptor-tyrosine kinase inhibitor resistance in non-small cell lung cancer. However, the comprehensive understanding of the KRAS pathway following the failure of epidermal growth factor receptor-tyrosine kinase inhibitor therapy remains limited. METHODS: We conducted a retrospective evaluation of the next generation sequencing data from 323 patients with advanced non-small cell lung cancer and EGFR-activating mutations after experiencing progression with epidermal growth factor receptor-tyrosine kinase inhibitor therapy. Our analysis specifically focused on the acquired changes to the KRAS gene. RESULTS: Among the 323 patients with advanced non-small cell lung cancer and EGFR-activating mutations who experienced resistance to epidermal growth factor receptor-tyrosine kinase inhibitor therapy, 14 individuals (4.3%) developed resistance due to acquired KRAS alterations. Of these 14 patients, 10 cases (71.4%) were due to KRAS missense mutations, 1 case (7.2%) was due to KRAS gene fusion and 3 cases (21.4%) were due to KRAS amplification. Notably, we identified one newly demonstrated KRAS gene fusion (KRAS and LMNTD1), one KRAS G13D and one KRAS K117N. The emergence of acquired KRAS alterations was often accompanied by novel mutations and high tumor mutation burden, with TP53, CNKN2A, PIK3CA, MYC, STK11, CDK4, BRCA2 and ERBB2 being the most frequently observed concurrent mutations. The median progression-free survival and overall survival for the 14 patients were 5.2 and 7.3 months, respectively. Acquired KRAS missense variants were associated with significantly worse progression-free survival compared with other KRAS variant subtypes (P < 0.028). CONCLUSIONS: This study provides significant evidence of the role of acquired KRAS variants in the development of resistance to epidermal growth factor receptor-tyrosine kinase inhibitor therapy. Our results contribute to the growing body of knowledge on the mutational profiles associated with resistance to epidermal growth factor receptor-tyrosine kinase inhibitor treatment. Furthermore, our study highlights the KRAS gene change as a significant mechanism of resistance to epidermal growth factor receptor-tyrosine kinase inhibitor therapy.

Whole-genome sequencing suggests a role of MIF in the pathophysiology of TEMPI syndrome.

TEMPI syndrome (telangiectasias, elevated erythropoietin level and erythrocytosis, monoclonal gammopathy, perinephric fluid collections, and intrapulmonary shunting) is a newly defined multisystemic disease with its pathophysiology largely unknown. Here, we report the whole-genome sequencing (WGS) analysis on the tumor-normal paired cells from a patient with TEMPI syndrome. WGS revealed somatic nonsynonymous single-nucleotide variants, including SLC7A8, NRP2, and AQP7. Complex structural variants of chromosome 2 were found, particularly within regions where some putative oncogenes reside. Of potential clinical relevance, duplication of 22q11.23 was identified, and the expression of the located gene macrophage migration inhibitory factor (MIF) was significantly upregulated in 3 patients with TEMPI syndrome. Importantly, the level of serum MIF in one patient with TEMPI syndrome was significantly decreased in accordance with the downtrend of plasma cells, M-protein, hemoglobin, and erythropoietin and the improvement of telangiectasias, perinephric fluid collections, and intrapulmonary shunting after treatment with plasma cell-directed therapy. In conclusion, our study provides insights into the genomic landscape and suggests a role of MIF in the pathophysiology of TEMPI syndrome.

A DNA methylation-associated nomogram predicts the overall survival of osteosarcoma.

ABSTRACT: Numerous reports have demonstrated that DNA methylation may be underlying prognostic biomarkers of cancer. However, few studies indicated that DNA methylation was independent biomarker for osteosarcoma prognosis. We aimed to discover and validate a novel DNA methylation signature for prediction of osteosarcoma patients' overall survival (OS).The DNA methylation data of osteosarcoma patients was researched from The Cancer Genome Atlas (TCGA) database. Overall, 80 samples with 485,577 DNA methylation sites were enrolled in our study. The 80 samples were randomly allocated into training dataset (first two-thirds) and validation dataset (remaining one-third). Initially, the univariate Cox proportional hazard analysis was performed in the training dataset to determine methylation sites significantly (P < .05) relevant to osteosarcoma patients' OS as underlying indicators. Subsequently, the underlying indicators were employed to carry out the least absolute shrinkage and selection operator (LASSO) Cox regression analysis for further selecting the candidate methylation sites. Then, the selected candidate methylation sites were employed as covariates to perform multivariate Cox proportional hazard model for identifying the predictor of OS in osteosarcoma patients. The validation dataset was used to validate the predictive accuracy by receiver operating characteristic (ROC) analysis and Kaplan-Meier survival analysis.We discovered a 7-DNA methylation signature closely relevant to OS of osteosarcoma patients. AUC at 1, 3, 5 years in training dataset (0.951, 0.922, 0.925, respectively), testing dataset (0.952, 0.918, 0.925, respectively), and entire dataset (0.952, 0.968, 0.968, respectively). Suggesting high predictive values for OS of osteosarcoma patients. In addition, a methylation-associated nomogram suggested good predictive value and clinical application.We discovered and validated a novel 7-DNA methylation-associated nomogram for predicting OS of osteosarcoma patients.

Mitochondria-targeting fluorescent molecules for high efficiency cancer growth inhibition and imaging.

Fluorescent mitochondria-accumulating delocalized lipophilic cations (DLCs) for cancer therapy have drawn significant attention in the field of cancer theranostics. One of the most promising fluorescent DLCs, F16, can selectively trigger the apoptosis and necrosis of cancer cells, making it an attractive targeted theranostic drug candidate. However, it suffers from low clinical translation potential, largely due to its inefficient anti-cancer activity (IC50 in the µM range) and poorly understood structure-activity relationship (SAR). In this report, eleven indole-ring substituted F16 derivatives (F16s) were synthesized. Among these derivatives, 5BMF was identified as a highly effective theranostic agent, with in vitro studies showing a low IC50 of ∼50 nM (to H2228 cells) and high cancer to normal cell selectivity index of 225. In vivo studies revealed that tumors treated with 5BMF were significantly suppressed (almost no growth over the treatment period) compared to the PBS treated control group, and also no obvious toxicity to mice was found. In addition, the tumor imaging capability of 5BMF was demonstrated by in vivo fluorescence imaging. Finally, we report for the first time a proposed SAR for F16 DLCs. Our work lays down a solid foundation for translating 5BMF into a novel and highly promising DLC for cancer theranostics.

Dual T1 and T2 weighted magnetic resonance imaging based on Gd3+ loaded bioinspired melanin dots.

In this report, a novel T1/T2 dual modal nanoprobe based on highly efficient and bioinspired melanin dots (M-dots) with directly loading gadolinium (Gd-M-dots) for magnetic resonance imaging (MRI) is described. In vitro and in vivo investigations have revealed that Gd-M-dots showed nontoxicity and good biocompatibilitity. Gd-M-dots relaxivity values on 3 T were determined to be r1 = 23.4 and r2 = 123.3 mM-1 s-1, which were much higher than both Gd-DTPA (r1 = 5.1, r2 = 6.2 mM-1 s-1) and Fe-M-dots (r1 = 1.2, r2 = 2.1 mM-1 s-1). For in vivo MRI, after injection of Gd-M-dots, simultaneous T1 and T2 contrast enhancement have been observed in the MRI of mice abdomen and mice bearing U87MG tumors. Furthermore, all the veins showed high signal intensity on T1-weighted MRI and remained for 2 h. Overall, in vitro and in vivo studies indicate that Gd-M-dot with high r1 relaxivity and r2 relaxivity has high potential to be a promising nanoprobe for MR venography and molecular imaging.

Chelator-Free and Biocompatible Melanin Nanoplatform with Facile-Loading Gadolinium and Copper-64 for Bioimaging.

Development of a chelator-free and biocompatible platform for the facile construction of gadolinium3+ (Gd3+)-loaded nanoparticle based probes for in vivo magentic resonance imaging (MRI) is still challenging. Herein, biocompatible Gd3+-loading melanin dots (Gd-M-dots) have been easily prepared and have exhibited good loading efficiency for Gd3+, high stability, and higher T1 relaxivity compared to the commercial Gd-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) agent. Furthermore, Gd-M-dots showed unique photoacoustic (PA) properties, and a high PA imaging signal could be observed in vivo 1 h after injection. Compared to the traditional Gd3+-loaded nanoparticles for single-modal MRI, Gd-M-dots can also be radiolabeled with 64Cu2+ for positron emission tomography. Overall, these attractive properties of Gd-M-dots render them a promising imaging agent for various biomedical applications.

Improved positron emission tomography imaging of glioblastoma cancer using novel 68Ga-labeled peptides targeting the urokinase-type plasminogen activator receptor (uPAR).

The urokinase-type plasminogen activator receptor (uPAR) is overexpressed in several cancers including glioblastoma (GBM) and is an established biomarker for metastatic potential. The uPAR-targeting peptide AE105-NH2 (Ac-Asp-Cha-Phe-(D)Ser-(D)Arg-Tyr-Leu-Trp-Ser-CONH2) is a promising candidate for non-invasive positron emission tomography (PET) imaging of uPAR. Despite the optimal physical properties of 68Ga for peptide-based PET imaging, low tumor uptakes have previously been reported using 68Ga-labeled AE105-NH2-based tracers. In an attempt to optimize the tumor uptake, we developed three novel tracers with alkane (AOC) and polyethylene glycol (PEG) spacers inserted between AE105-NH2 and the radio metal chelator 2-(4,7-bis(carboxymethyl)-1,4,7-triazonan-1-yl)pentanedioic acid (NODAGA). The resulting tracers NODAGA-AOC-AE105-NH2, NODAGA-PEG3-AE105-NH2 and NODAGA-PEG8-AE105-NH2 were compared to the non-spacer version, NODAGA-AE105-NH2. Following radiolabeling with 68Ga, we evaluated the in vitro and in vivo performance in mice bearing subcutaneous tumors derived from the uPAR-expressing human GBM cell line U87MG. In vivo PET/CT imaging showed that introduction of PEG spacers more than doubled the in vivo tumor uptake after 1 h compared with the non-spacer version: 68Ga-NODAGA-PEG3-AE105-NH2 (2.08 ± 0.37%ID/g) and 68Ga-NODAGA-PEG8-AE105-NH2 (2.01 ± 0.22%ID/g) vs. 68Ga-NODAGA-AE105-NH2 (0.70 ± 0.40%ID/g), p < 0.05. In addition, 68Ga-NODAGA-PEG8-AE105-NH2 showed significantly higher (p < 0.05) tumor-to-background contrast (3.68 ± 0.23) than the other tracers. The specific tumor-targeting property of 68Ga-NODAGA-PEG8-AE105-NH2 was established by effectively blocking the tumor uptake with co-injection of unlabeled AE105-NH2 (1 h: unblocked 2.01 ± 0.22%ID/g vs. blocked 1.24 ± 0.09%ID/g, p < 0.05). Ex vivo biodistribution confirmed the improved tumor uptakes of the PEG-modified tracers. 68Ga-NODAGA-PEG8-AE105-NH2 is thus a promising candidate for human translation for PET imaging of GBM.

The clinical usefulness of (99m)Tc-Tetrofosmin scintigraphy in the diagnosis of lung neoplasmas and mediastinal lymphoid node involvement.

In order to investigate the clinical significance of (99m)Tc-Tetrofosmin (TF) scintigraphy in the evaluation of lung cancer and mediastinal lymphoid node involvement, 33 patients with pulmonary neoplasmas were subjected to both (99m)Tc-TF scintigraphies and CT scans in one week before their operations or puncturations. All the images were judged visually and the emission images were analyzed with semi-quantitative methods in addition. The results of each group were compared. There was marked difference in target/non-target (T/N) ratio between the lung cancer group and the benign lesion group (P<0.001). Moreover, in the lung cancer group, T/N ratio in tomographies was significantly higher than that in planar images (P<0.01). The sensitivity and accuracy of semi-quantitative analysis in (99m)Tc-TF SPECT were significantly higher than those of CT in the diagnosis of pulmonary neoplasmas (P<0.05 and P<0.01 respectively), so was the sensitivity of (99m)Tc-TF SPECT vs CT in the diagnosis of mediastinal lymphoid node metastasis (P<0.05). It was also found that epidermoid squamous cell carcinomas and adenocarcinomas had a higher T/N ratio than in small cell carcinomas (P<0.05), and 2 h washout rate (WR) of adenocarcinomas was higher than that of epidermoid squamous cell carcinomas (P<0.05). In conclusion, (99m)Tc-TF scintigraphy showed a favorable diagnostic accuracy in appraising lung cancers and mediastinal lymph node metastases. Furthermore semi-quantitative technology can improve the accuracy, and is potential to offer some information about histological type of the cancer tissue. Therefore, (99m)Tc-TF scintigraphy will be a useful tool in the diagnosis and staging of lung cancer.

[Clinical significance of 99mTc-tetrofosmin SPECT in the diagnosis of lung neoplasms and mediastinal lymph node involvement].

BACKGROUND: An accurate pre-operative tumor staging is the premise for a successful operation, for which a non-invasive evaluation of the nature of lung nodules and the real extent of them is necessary. As the pioneer of functional images, nuclear medicine continues to introduce novel radiopharmaceuticals and new techniques for cancer imaging, including tetrofosmin (TF). The aim of this study is to evaluate the clinical usefulness of 99m Tc-tetrofosmin SPECT ( 99m Tc-TF SPECT) in the diagnosis of lung cancer and mediastinal lymph node involvement. METHODS: Thirty-three patients with pulmonary neoplasms took both CT scan and 99m Tc-TF SPECT in one week before their operations or puncture biopsy. Then the sensitivity, specificity and accuracy were acquired according to the pathological outcomings and the results of 99m Tc-TF SPECT and CT were compared in detection of lung cancer and mediastinal lymph node metastasis, respectively. RESULTS: The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of 99m Tc-TF SPECT for lung cancer were 95.7%, 80.0%, 90.9%, 91.7%, 88.9%, and 73.9%, 70.0%, 72.7% , 85.0% , 53.8% by CT, respectively. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of 99m Tc-TF SPECT for mediastinal lymph node metastasis were 93.3%, 75.0%, 87.0% , 87.5%, 85.7%, while those of CT were 60.0%, 75.0%, 65.2%, 81.8%, 50.0% respectively. The sensitivity of 99m Tc-TF SPECT was significantly higher than that of CT for lung neoplasms (P < 0.05). CONCLUSIONS: 99m Tc-TF SPECT is a useful non-invasive presurgical tool to evaluate primary lung cancers and mediastinal lymph node involvement, and it will play a favorable clinical role in diagnosis and staging of lung cancer.