Baoming Mi

Background Cisplatin-based concurrent chemotherapy is considered first-line treatment for advanced NSCLC. In this study, the role of ⁶⁸Ga-FAPI in visualizing chemotherapy resistance and the regulatory mechanisms which CAFs influence chemoresistance of NSCLC was evaluated. Methods The CAFs were isolated form fresh NSCLC tissue, and the expression of FAP was evaluated by qRT-PCR and western blotting. ⁶⁸Ga-FAPI micro-PET/CT was performed to visualize CAFs distribution and quantity in vivo. A cytokine array was conducted to analyze the secretion of HGF. The expression of LINC01123 was detected by overlapping high-throughput sequencing results with the analysis of the GEO database (GSE43493). Finally, the interaction between LINC01123 and β-catenin was assessed using RNA pull-down and RIP techniques. Results In this study, we employed ⁶⁸Ga-FAPI micro-PET/CT to quantify the number of CAFs and visualize the different uptake between cisplatin sensitive and resistant NSCLC xenograft models. The biological role of CAFs in cisplatin treatment for NSCLC was evaluated through functional experiments in vitro and in vivo. Functional assay demonstrated that CAFs secrete HGF which upregulates the expression of LINC01123 in NSCLC cells. LINC01123 directly binds to β-catenin and enhances its transcription activity. The activated HGF/LILNC01123/β-catenin signaling-mediated crosstalk between CAFs and tumor cells drives the chemoresistance of NSCLC. Conclusions Dysregulated activation of the HGF/LINC01123/β-catenin cascade represents a pivotal pathway through which CAFs interact with NSCLC cells, which enhancing the role of ⁶⁸Ga-FAPI to visualize chemotherapy resistance in patients.

Objective: Overexpression of human epidermal growth factor receptor-2 (HER2) in breast cancers provides promising opportunities for imaging and targeted therapy. Developing HER2 targeted positron emission tomography (PET) probes might be benefit for management of the disease. Small high-affinity scaffold proteins, affibodies, are ideal vectors for imaging HER2 overexpressed tumors. Despite of the initial success on development of 18F labeled ZHER2:342 affibody, the tedious synthesis producers, low yields and unfavorable pharmacokinetics may hinder the clinical use. 68Ga is an attractive positron emitter for PET imaging. A simple preparation of 68Ga labeled ZHER2:342 analog, 68Ga-NOTA-MAL-Cys-MZHER2:342, was reported in the study. The in vivo performances of the tracer for assessing HER2 status in breast cancers were also evaluated. Methods: NOTA-MAL conjugated Cys-MZHER2:342 was radiolabeled with 68Ga. The probe was evaluated by in vitro tests including stability and cell binding studies in breast cancer cells with different HER2 levels. In vivo evaluation was performed in mice bearing tumors using microPET imaging and biodistribution experiments. A PET/CT imaging study was initially performed in patients with breast cancers. Results: The tracer was synthesized in a straightforward chelation method with satisfactory non-decay corrected yield (81±5%) and radiochemical purity (>95%). In vivo micro-PET imaging showed that HER2 high levels expressed BT474 xenografts were more clear visualized than HER2 low levels expressed MCF-7 tumors (16.12 ± 2.69 ID%/g vs 1.32 ± 0.19 ID%/g at 1 h post-injection). The outcome was consistent with the immunohistochemical analysis. No significant radioactivity was accumulated in healthy tissues (less than 2% ID/g) except kidneys. In a preliminary clinical study, 68Ga-NOTA-MAL-Cys-MZHER2:342 PET imaging allowed more high-contrast detection of HER2 positive primary tumors (maximum standardized uptake value = 2.16±0.27) than those in HER2 negative primary focus (maximum standardized uptake value = 0.32±0.05). No detectable side-effects were found. Conclusion: In summary, this study indicates the significant efficiency of the 68Ga labeled HER2 affibody. Preclinical and clinical studies support the possibility of monitoring HER2 levels in breast cancers using 68Ga-NOTA-MAL-Cys-MZHER2:342. Advances in knowledge: The research investigated the feasibility of a 68Ga labeled HER2 affibody modified with a hydrophilic linker for breast cancer PET imaging. Favorable outcomes showed that the probe might be valuable for determining HER2 status of the disease.

Background: Long non-coding RNAs (lncRNAs) have been associated with non-small cell lung cancer (NSCLC), but the underlying molecular mechanisms of their specific roles in mediating aerobic glycolysis have been poorly explored. Methods: Next-generation RNA sequencing assay was performed to identify the differentially expressed RNAs between NSCLC tissues with high ¹⁸F-fluorodeoxyglucose (FDG) uptake and their adjacent normal lung tissues. LINC01123 expression in NSCLC tissues was measured by real-time PCR and in situ hybridization (ISH) assay. The biological role of LINC01123 in cell growth and aerobic glycolysis capability was determined by performing functional experiments in vitro and in vivo. Further, the transcription of LINC01123 was explored by bioinformatics analysis, dual-luciferase reporter assay, and chromatin immunoprecipitation (ChIP) assay. RNA immunoprecipitation (RIP) and luciferase analyses were used to confirm the predicted competitive endogenous RNA (ceRNA) mechanisms between LINC01123 and c-Myc. Results: Three hundred sixty-four differentially expressed genes were identified in RNA-seq assay, and LINC01123 was one of the most overexpressed lncRNAs. Further validation in expanded NSCLC cohorts confirmed that LINC01123 was upregulated in 92 paired NSCLC tissues and associated with poor survival. Functional assays showed that LINC01123 promoted NSCLC cell proliferation and aerobic glycolysis. Mechanistic investigations revealed that LINC01123 was a direct transcriptional target of c-Myc. Meanwhile, LINC01123 increased c-Myc mRNA expression by sponging miR-199a-5p. In addition, rescue experiments showed that LINC01123 functioned as an oncogene depending on miR-199a-5p and c-Myc. Conclusion: Since LINC01123 is upregulated in NSCLC, correlates with prognosis, and controls proliferation and aerobic glycolysis by a positive feedback loop with c-Myc, it is expected to be a potential biomarker and therapeutic target for NSCLC.