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

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.

Small cell lung cancer (SCLC) is known as recalcitrant cancer with high malignancy and heterogeneity. Immunotherapy has changed the treatment pattern of extensive-disease SCLC (ED-SCLC), but the beneficiary population is limited. Therefore, exploring new therapeutic strategies is an urgent clinical problem to be solved for SCLC. SCLC is characterized by highly active glycolytic metabolism and pyruvate kinase M1 (PKM1) is one of the isozymes of PK, an important rate-limiting enzyme in glycolysis pathway. Previous studies have shown that PKM1 is related to autophagy and drug sensitivity, however, how PKM1 regulates drug sensitivity in SCLC and its mechanism remain unclear. The aim of this study was to investigate the biological functions of PKM1 in SCLC, including its effects on proliferation, migration, autophagy, drug sensitivity, and expression of neuroendocrine (NE)-related markers in SCLC.

[PKM1 Regulates the Expression of Autophagy and Neuroendocrine Markers  in Small Cell Lung Cancer].

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.

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 KRAS mutation represents the most prevalent oncogenic alteration observed in human cancers. Its primary role involves directly driving malignant tumor development and growth through the activation of downstream signaling pathways. Increasing evidence suggests that KRAS significantly affects the immune response of KRAS-mutant tumors by modulating immune-related signaling and inflammatory pathways. In addition to broadly regulating the KRAS-associated immune signaling, KRAS influences immune cell phenotype and function by triggering tumor metabolic pathways. Here, we reviewed the KRAS mutation-associated immune microenvironment features and discussed how KRAS remodels the immune microenvironment by regulating immune-related molecules, inflammatory factors, and multiple metabolic pathways, offering insights that could be useful for developing effective immune-responsive therapies for KRAS-mutant tumors.

Article	Year
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
[PKM1 Regulates the Expression of Autophagy and Neuroendocrine Markers  in Small Cell Lung Cancer]. Zhongguo fei ai za zhi = Chinese journal of lung cancer\| PubMed ID: 39492579	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
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
Oncogenic KRAS mutations drive immune suppression through immune-related regulatory network and metabolic reprogramming. Cell death & disease\| PubMed ID: 41184283	2025

Chenchen Tang

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