effects-on-rna-quality-pbmcs-under-different-cryopreservation-times

Effects on RNA Quality of PBMCs Under Different Cryopreservation Times and Processing Methods in Patients with Advanced Non-Small Cell Lung Cancer

Cryopreserved peripheral blood mononuclear cells (PBMCs) are widely used in RNA sequencing (RNA-seq), and the quality of their RNA directly influences ...

Jilin Cancer Hospital

 To investigate the association between serum lipid levels in patients with primary non-small cell lung cancer and the risk of developing metastases, a retrospective cohort-based nested case-control study was conducted.  Patients with primary non-small cell lung cancer admitted to the First and the Third Hospitals of Jilin University from January 2008 through December 2015 were recruited retrospectively based on their electronic medical records. A total of 524 patients were initially considered, consisting of 138 in the case group and 386 as control. Out of these, 110 were finally included in the case group and 110 as control based on additional selection criteria. The following information is collected from all the patients, total cholesterol (TC), low-density lipoprotein (LDL-C), high density lipoprotein (HDL-C) and triglyceride (TG). Logistic regressions were conducted to estimate the odds ratios (ORs) and 95% confidence intervals (95% CI) for non-small cell lung cancer (NSCLC) patients to have metastasis risk when having elevated serum lipid levels. Restricted cubic spline (RCS) curves were used to demonstrate the association between serum lipid levels and the risk of metastasis.  Patients with high TC level ( = 0.025, 0R = 1.35, 95% CI: 1.03-1.74) and patients with high LDL-C level (Q4: > 3.47 vs Q1: ≤2.54,  = 0.002, OR = 3.92, 95% CI: 1.31-11.77) are found to have an increased metastasis risk; and their dose-response relationship was validated by our restricted cubic spline analysis (TC:  overall association=0.02, P non-linear association = 0.73; LDL-C: P overall association=0.02,  non-linear association = 0.10). These associations were statistically significant, particularly in men who smoked, never drank, and were 65 years of age or younger. In addition, patients with simultaneously high levels of TC and LDL-C have a 60% increased risk of metastasis compared with patients with high levels of TC and normal LDL-C.  Dyslipidemia may be a risk factor for metastasis among NSCLC patients. Examination of serum lipid level on a regular basis can provide early signal of metastasis for NSCLC patients.

Elevated serum lipid level can serve as early signal for metastasis for Non-Small Cell Lung Cancer patients: A retrospective nested case-control study.

 mutation is one of important driver genes in non-small-cell lung cancer (NSCLC) and the patients with   mutations benefit from the inhibitor AMG510. However, the frequency, concurrent pathogenic mutations, and clinical characteristic of   is unknown in the NSCLC population of Northeast China.

The Prevalence and Concurrent Pathogenic Mutations of   in Northeast Chinese Non-small-cell Lung Cancer Patients.

Relapse and drug resistance are the main causes of mortality in patients with small‑cell lung cancer (SCLC). Intratumoral heterogeneity (ITH) is a key biological mechanism that leads to relapse and drug resistance. Phenotypic plasticity is an important factor that leads to ITH in SCLC, although its mechanisms and key regulatory factors remain to be elucidated. In the present study, cell proliferation and cell switch assay were measured using trypan blue. Alamar Blue was used to test drug sensitivity. Differential genes were screened by RNA sequencing. Reverse transcription‑quantitative PCR and western blotting were performed to assess the expressions of CSF2/p‑STAT3/MYC pathway related molecules, neuroendocrine (NE)/non‑neuroendocrine (non‑NE), transcription factors and drug‑related targets. The present study found that SCLC cell line NCI‑H69 exhibited adherent (H69A) and suspensive (H69S) phenotypes, which could switch back and forth. The two phenotypic cells had significant differences in cellular NE and non‑NE characteristics, drug sensitivity and expression of drug‑related targets. RNA sequencing showed that granulocyte‑macrophage colony‑stimulating factor [i.e., colony‑stimulating factor 2 (CSF2)] was the main differentially expressed gene between the two phenotypes and that H69A cells highly expressed CSF2. The inhibition of CSF2 promoted the transformation from H69A to H69S, increased drug sensitivity and NE marker expression and decreased the non‑NE marker expression in H69A. The STRING, Pathway Commons and Reactome databases showed a potential regulatory relationship between CSF2 and phosphorylated signal transducer and activator of transcription 3 (p‑STAT3)/MYC. p‑STAT3 and MYC expression was higher in H69A cells than in H69S cells and CSF2 silencing inhibited their expression. Taken together, these results indicated that CSF2 may regulate the phenotypic plasticity of SCLC through the phosphorylated STAT3/MYC pathway, thereby limiting the transformation between cell clones with different phenotypes and changing the sensitivity of specific cell clones to targeted drugs. Targeting CSF2 may be a potential therapeutic strategy to overcome drug resistance in SCLC treatment by influencing ITH.

Colony‑stimulating factor CSF2 mediates the phenotypic plasticity of small‑cell lung cancer by regulating the p‑STAT3/MYC pathway.

Lung adenocarcinoma (LUAD) is the most common pathological type of lung cancer. In recent years, immunotherapy has greatly changed the treatment pattern of advanced LUAD. However, only a small proportion of LUAD patients benefitted from immune checkpoint inhibitor therapy. There is an urgent need to develop a biomarker to predict immune therapy response. E2F7 has been shown to be closely related to immune cell infiltration and immune checkpoint expression in tumors. However, it is unclear whether the E2F7 expression is related to the immunotherapy efficacy in LUAD. Therefore, we conducted this study to investigate the clinical characteristics, function, and immunotherapy responsiveness of E2F7 expression, and to explore the potential of E2F7 as an immunotherapy response biomarker in LUAD. We analyzed the clinical characteristics and biological function of E2F7 expression based on data from the Cancer Genome Atlas and Gene Expression Omnibus database. In addition, we used single-cell sequencing data to analyze the immune regulatory effects of E2F7 in LUAD. Furthermore, we analyzed the immunotherapy response prediction ability of E2F7 expression based on the immunotherapy database. Compared to normal lung tissue, E2F7 was specifically overexpressed in LUAD, and its expression was associated with higher malignancy and poor efficacy. E2F7 high expression was an independent risk factor affecting the prognosis of LUAD. E2F7 was enriched in cell division and cell cycle functions. In addition, the expressions of immune checkpoints were correlated with the E2F7 expression. E2F7 was highly expressed in myeloid cells, and E2F7 highly expressed myeloid cells were associated with immune and inflammatory responses. Moreover, the expression level of E2F7 can effectively distinguish different immune therapy responses in LUAD patients. E2F7 was upregulated in LUAD, and high expression of E2F7 was associated with higher malignancy and poor efficacy. E2F7 high expression was an independent risk factor affecting the prognosis of LUAD. Moreover, E2F7 may exert its immunosuppressive effect by affecting the function of myeloid cells. These results indicated the potential role of E2F7 as a biomarker for predicting LUAD immunotherapy responses.

The predictive value of E2F7 in immunotherapy efficacy for lung adenocarcinoma: An observational study.

Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha   mutations are associated with drug resistance and prognosis in lung cancer; however, the consistency and clinical value of  mutations between tissue and liquid samples are unknown.

Highly consistency of  mutation spectrum between circulating tumor DNA and paired tissue in lung cancer patients.

There is an urgent need to develop new targeted treatment agents for small cell lung cancer (SCLC). Tinengotinib (TT-00420) is a novel, multi-targeted, and spectrally selective small-molecule kinase inhibitor that has shown significant inhibitory effects on certain solid tumors in preclinical studies. However, its role and mechanism of action in SCLC remain unclear. In this study, we demonstrated that tinengotinib effectively inhibited SCLC cell proliferation, especially highly expressing NeuroD1 (SCLC-N), in the SCLC cell line-derived xenograft (CDX) model and the malignant pleural effusion cell model of patients with SCLC. When combined with etoposide/cisplatin, it synergistically inhibited SCLC growth. Tinengotinib regulates proliferation, apoptosis, migration, cell cycle and angiogenesis in SCLC cells. Mechanistic studies revealed that c-Myc expression may be a key factor influencing the effect of tinengotinib in SCLC-N. This study provides reliable preclinical data and a new direction for tinengotinib as a promising therapy for SCLC, either alone or in combination with chemotherapy.

Multiple Kinase Small Molecule Inhibitor Tinengotinib (TT-00420) Alone or With Chemotherapy Inhibit the Growth of SCLC.

Small cell lung cancer (SCLC) is acknowledged as the most aggressive histological subtype among the various types of lung cancer. Among patients with advanced-stage SCLC, liver metastasis (LM) is frequently observed and is associated with the most unfavorable prognosis. The purpose of the present study is to investigate the synchronous metastatic sites in SCLC patients with LM, and to construct a nomogram for predicting the survival of these patients.

Metastatic sites, prognosis, and a new nomogram for predicting overall survival among small cell lung cancer patients with liver metastasis: a retrospective study based on SEER.

Observational studies have shown that gallstone disease (GSD), cholecystitis, cholangitis, polyp of gallbladder, viral hepatitis, pancreatitis and gastrointestinal (GI) traits such as H. pylori infection, inflammatory bowel disease, and digestive ulcer are associated with the risk of biliary tract cancer (BTC). However, no study has explored their causal associations.

Gastrointestinal traits, common inflammatory disorders, gallstones, and biliary tract cancer: A network Mendelian randomization study.

Tyrosine kinase inhibitor (TKI)-targeted therapy has significantly improved the quality life of patients with advanced non-small cell lung cancer (NSCLC). Smoking is the main pathogenic factor of lung cancer. A number of the studies have analyzed smoking patients and nonsmoking patients as grouping factors, but few have examined the characteristic gene mutations in these groups. This study thus aimed to examine the characteristic pathogenic mutations in smokers with NSCLC.

The pathogenic mutations in smokers with non-small cell lung cancer: a next-generation sequencing study.

Conventional three-dimensional (3D) organoid models are limited by high cost, technical complexity, and poor replication of tumor-immune interactions. To address these limitations, we developed two novel 2.5D systems from malignant pleural effusion (MPE) of patients with lung adenocarcinoma. The first uses MPE-derived adherent cells to create matrix-free niches, while the second combines MPE supernatant with DMEM-F12 and 10-30 % fetal bovine serum, with or without agarose, to form a thermoresponsive matrix that mimics dynamic tumor microenvironments. Autologous MPE supernatants were used to preserve native cytokines and extracellular matrix components. Genomic and immune microenvironment analyses demonstrated over 90 % concordance between 2.5D organoids and parental tumors, which aligned with the standards reported for existing 3D tumor-organoid cultures. The 2.5D organoids retained key functional characteristics, including mucus secretion, phenotypic plasticity, and angiogenesis. Organoid-based high-throughput screening enabled identification of candidate therapeutics, and drug sensitivity assays correlated strongly with patient outcomes, underscoring their utility for personalized medicine. The matrix-free design facilitated rapid organoid generation (3-10 days) and reduced costs by approximately 90 % relative to conventional 3D methods. These 2.5D models provide a clinically translatable platform that replicates tumor heterogeneity while overcoming cost and technical barriers, offering a promising tool for precision pharmacology and oncology.

A clinically translatable 2.5D lung adenocarcinoma organoid platform derived from malignant pleural effusions for precision drug screening and tumor-immune-stromal modeling.

The treatment of patients with advanced epidermal growth factor receptor (EGFR)-mutated non-small-cell lung cancer (NSCLC) whose disease progresses after tyrosine-kinase inhibitors (TKIs) treatment has become a research hotspot.

Efficacy and safety analysis of treatment in patients with EGFR-mutated advanced NSCLC who progressed on TKIs: a systematic review and meta-analysis.

To investigate the clinical characteristics and impact of  mutations in patients with non-small cell lung cancer (NSCLC).

Clinicopathological characteristics and therapeutic outcomes in patients with non-small cell lung cancer harboring  mutations.

For limited-stage small cell lung cancer (LS-SCLC), immunotherapy combined with chemotherapy as induction followed by subsequent radiotherapy is still under clinical investigation. Some patients who receive treatment have undergone radiotherapy following immunochemotherapy in real-world. However, the preliminary efficacy and safety remain unknown. In this study, we will explore a novel treatment approach for LS-SCLC.

Article	Year
Elevated serum lipid level can serve as early signal for metastasis for Non-Small Cell Lung Cancer patients: A retrospective nested case-control study. Journal of Cancer\| PubMed ID: 33123292	2020
The Prevalence and Concurrent Pathogenic Mutations of in Northeast Chinese Non-small-cell Lung Cancer Patients. Cancer management and research\| PubMed ID: 33758543	2021
Colony‑stimulating factor CSF2 mediates the phenotypic plasticity of small‑cell lung cancer by regulating the p‑STAT3/MYC pathway. Oncology reports\| PubMed ID: 35583004	2022
The predictive value of E2F7 in immunotherapy efficacy for lung adenocarcinoma: An observational study. Medicine\| PubMed ID: 38905387	2024
Highly consistency of mutation spectrum between circulating tumor DNA and paired tissue in lung cancer patients. Heliyon\| PubMed ID: 39071569	2024
Multiple Kinase Small Molecule Inhibitor Tinengotinib (TT-00420) Alone or With Chemotherapy Inhibit the Growth of SCLC. Cancer science\| PubMed ID: 39817471	2025
Metastatic sites, prognosis, and a new nomogram for predicting overall survival among small cell lung cancer patients with liver metastasis: a retrospective study based on SEER. Journal of thoracic disease\| PubMed ID: 39975725	2025
Gastrointestinal traits, common inflammatory disorders, gallstones, and biliary tract cancer: A network Mendelian randomization study. Journal of advanced research\| PubMed ID: 40064441	2026
The pathogenic mutations in smokers with non-small cell lung cancer: a next-generation sequencing study. Translational lung cancer research\| PubMed ID: 41132982	2025
A clinically translatable 2.5D lung adenocarcinoma organoid platform derived from malignant pleural effusions for precision drug screening and tumor-immune-stromal modeling. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie\| PubMed ID: 41175595	2025
Efficacy and safety analysis of treatment in patients with EGFR-mutated advanced NSCLC who progressed on TKIs: a systematic review and meta-analysis. Frontiers in immunology\| PubMed ID: 41229435	2025
Clinicopathological characteristics and therapeutic outcomes in patients with non-small cell lung cancer harboring mutations. Annals of medicine\| PubMed ID: 41593906	2026
Real-world evidence on the efficacy, safety, and biomarkers of immunotherapy combined with radiotherapy in limited-stage small cell lung cancer. Journal of thoracic disease\| PubMed ID: 41816449	2026

Rixin Li

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