Abstract Background Immune‐therapy with anti‐PD1 inhibitors, such as pembrolizumab, is revolutionizing the treatment of non‐small cell lung cancers (NSCLC). However, identifying patients for the potential therapeutic response and predicting therapy resistance and early relapse remains a challenge. Methods Between 2016 and 2018, 60 patients were treated with pembrolizumab, among who 12 NSCLC patients had both baseline (before treatment) and serial (on treatment) periodical circulating tumor DNA (ctDNA) samples. Those samples were sequenced on a 329 pan cancer‐related gene panel. Analyses of tumor burden, blood tumor mutational burden (bTMB), maximum somatic allele frequency (MSAF), and tumor clonal structure were performed in association with clinical response. Candidate resistance mutations involved in relapse and metastases were further investigated. Results ctDNA was detected and mutational profiling was performed for each patient. Those with a high baseline bTMB level showed significantly improved progression‐free survival (PFS) after pembrolizumab treatment. Tumor burden and therapeutic response significantly correlated with the MSAF instead of the bTMB. Clone analysis detected tumor progression about 2‐4 months ahead of computed tomography (CT) scan. One mutation in gene PTCH1 (Protein patched homolog 1) and two acquired anti‐PD1 candidate resistance mutations of gene B2M (β2 microglobulin) were identified in association with distant metastasis. The evolutionary tree of a representative patient was also described. Conclusion This pilot study showed that MSAF could be another good indicator of therapeutic response, and clonal analysis could be clinically useful in monitoring clonal dynamics and detecting remote metastasis and early relapse.
Two Autographa californica nucleopolyhedrovirus (AcMNPV) encoded miRNAs, AcMNPV-miR-1 and AcMNPV-miR-3, have been reported by us in 2013 and 2019, respectively. Here, we present an integrated investigation of AcMNPV-encoded miRNAs, which include the above two miRNAs and three additional newly identified miRNAs. Six candidate miRNAs were predicted through small RNA deep sequencing and bioinformatics, of which, five were validated. Three miRNAs are located opposite the coding sequences, the other two are located in the coding sequences of viral genes. Targets in both virus and host were predicted and subsequently tested using dual-luciferase reporter assays. The validated targets were found mainly in AcMNPV, except for the targets of AcMNPV-miR-4, which are all host genes. Based on reporter assays, the five miRNAs predominantly function by down-regulating their targets. The transcription start sites of these miRNAs were bioinformatic screened based on known baculovirus promoter motifs. Our study reveals that AcMNPV-encoded miRNAs function as fine modulators of the interactions between host and virus by regulating viral and/or host genes.
The purpose of the experiment was mainly to establish and verify a precise and straightforward ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach for simultaneously analyzing the concentration levels of amiodarone, dronedarone, and their metabolites (desethylamiodarone and desbutyldronedarone) in the plasma of Sprague-Dawley (SD) rats and to investigate the pharmacokinetics of all analytes in SD rats. After rapid protein precipitation by using acetonitrile, we accomplished the chromatographic separation of amiodarone, desethylamiodarone, dronedarone, desbutyldronedarone and ivabradine (internal standard, IS) by using Acquity BEH C18 column and detected through a mass spectrometer with Xevo TQ-S triple quadrupole tandem, choosing the positive ion mode. The approach showed wonderful linearity, and the range of calibration curve for amiodarone was 1–200 ng/mL, desethylamiodarone was 0.1–20 ng/mL, dronedarone was 0.5–100 ng/mL, and desbutyldronedarone was 0.25–50 ng/mL, respectively. For lower limit of quantification (LLOQ), the current method of UPLC-MS/MS can achieve values of 1.0 ng/mL for amiodarone, 0.1 ng/mL for desethylamiodarone, 0.5 ng/mL for dronedarone, and 0.25 ng/mL for desbutyldronedarone, respectively. The accuracy of intra-day and inter-day of all analytes was between −14.8% to 10.9%, while the precision was ≤ 13.3%. For each substance, the recovery rate was > 82.1%, besides, obvious matrix effect was not found. In all conditions, the stability of all analytes was comfirmed to the plasma sample quantification. In addition, the method of UPLC-MS/MS we developed could also be applied to measure the pharmacokinetic characteristics including amiodarone, desethylamiodarone, dronedarone, and desbutyldronedarone in the plasma of SD rats.
Background: MET amplification is one of the EGFR-independent mechanisms of EGFR tyrosine kinase inhibitor (TKI) resistance. Combinatorial therapy of EGFR-TKI and crizotinib has been explored as a strategy to overcome resistance by simultaneously targeting both EGFR and MET pathways; however, no consensus still exists on the optimal combination regimen with the most benefit.Methods: Retrospective analysis was performed on the clinical and sequencing data obtained from eleven patients with lung adenocarcinoma who acquired MET amplification at progression from prior EGFR-TKI therapy and received a combination of EGFR-TKI and crizotinib.Results: Acquired MET amplification was detected in four and seven patients who progressed from first-line gefitinib and second-line osimertinib, respectively. Six and five patients received a combination of either first-generation (gefitinib, erlotinib, or icotinib) or third-generation (osimertinib) EGFR-TKI and crizotinib, respectively. Nine patients achieved partial response, resulting in an overall response rate of 81.8 %. The median progression-free survival of the cohort was 5.8 months. Moreover, analysis of acquired resistance mechanisms from nine patients identified EGFR T790 M from three patients who progressed from first-generation EGFR-TKI and crizotinib, while EGFR T790 M/trans-C797S and L718Q, EGFR G724S, and CCDC6-RET fusion were detected from one patient each who progressed from osimertinib and crizotinib regimen. Loss of MET amplification was also observed in a majority of the patients at progression from the combination therapy.Conclusions: Our study provides clinical evidence of the efficacy of combinatorial regimen with either first- or third-generation EGFR-TKI and crizotinib after the emergence of MET amplification-mediated EGFR-TKI resistance in patients with EGFR-mutant NSCLC.
To further our understanding of the pathobiology of esophageal squamous cell carcinoma (ESCC), we previously performed microRNA profiling that revealed downregulation of miR-200b in ESCC. Using quantitative real-time PCR applied to 88 patient samples, we confirmed that ESCC tumors expressed significantly lower levels of miR-200b compared with the respective adjacent benign tissues ( P = 0.003). Importantly, downregulation of miR-200b significantly correlated with shortened survival ( P = 0.025), lymph node metastasis ( P = 0.002) and advanced clinical stage ( P = 0.020) in ESCC patients. Quantitative mass spectrometry identified 57 putative miR-200b targets, including Kindlin-2, previously implicated in the regulation of tumor invasiveness and actin cytoskeleton in other cell types. Enforced expression of miR-200b mimic in ESCC cells led to a decrease of Kindlin-2 expression, whereas transfection of miR-200b inhibitor induced Kindlin-2 expression. Furthermore, transfection of miR-200b mimic or knockdown of Kindlin-2 in ESCC cells decreased cell protrusion and focal adhesion (FA) formation, reduced cell spreading and invasiveness/migration. Enforced expression of Kindlin-2 largely abrogated the inhibitory effects of miR-200b on ESCC cell invasiveness. Mechanistic studies revealed that Rho-family guanosine triphosphatases and FA kinase mediated the biological effects of the miR-200b—Kindlin-2 axis in ESCC cells. To conclude, loss of miR-200b, a frequent biochemical defect in ESCC, correlates with aggressive clinical features. The tumor suppressor effects of miR-200b may be due to its suppression of Kindlin-2, a novel target of miR-200b that modulates actin cytoskeleton, FA formation and the migratory/invasiveness properties of ESCC.
The clinical outcome of resectable non-small-cell lung cancer (NSCLC) patients receiving neoadjuvant chemoimmunotherapy is good but varies greatly. In addition, the pathological response after neoadjuvant chemoimmunotherapy is significantly associated with survival outcomes. The aim of this retrospective study was to identify which population of patients with locally advanced and oligometastatic NSCLC has a favorable pathological response after neoadjuvant chemoimmunotherapy. NSCLC patients treated with neoadjuvant chemoimmunotherapy were enrolled between February 2018 and April 2022. Data on clinicopathological features were collected and evaluated. Multiplex immunofluorescence was performed on pre-treatment puncture specimens and surgically resected specimens. In total, 29 patients with stages III and IV locally advanced or oligometastatic NSCLC who received neoadjuvant chemoimmunotherapy and R0 resection were enrolled. The results showed that 55% (16/29) of patients had a major pathological response (MPR) and 41% (12/29) of patients had a complete pathological response (pCR). In the stroma area of the pre-treatment specimen, the higher infiltration of CD3
Studies have shown that functional abnormalities in the locus coeruleus (LC) are strongly associated with depressive symptoms, but the pattern of LC functional connectivity in Alzheimer's disease patients with depressive symptoms (D-AD) remains unclear. The current study aimed to investigate the characteristics of LC functional connectivity (FC) in D-AD using resting-state functional magnetic resonance imaging (rsfMRI). We obtained rsfMRI data in 24 D-AD patients (aged 66–76 years), 14 non-depressive AD patients (nD-AD) (aged 69–79 years) and 20 normal controls (aged 67–74 years) using a 3 T scanner. We used the FC approach to investigate abnormalities in the LC brain network of D-AD patients. One-way ANCOVA and post-hoc two-sample t-tests were performed to compare the strength of functional connectivity from the LC among the three groups. Our results showed that, compared with normal controls, D-AD showed decreased left LC FC with the right caudate and left fusiform gyrus, whereas nD-AD showed decreased left LC FC with the right caudate, right middle frontal gyrus and left fusiform gyrus. Compared to nD-AD, D-AD showed increased left LC FC with right superior frontal gyrus and right precentral gyrus. These findings contribute to our understanding of the neural mechanisms of D-AD.
ABSTRACT Baculovirus-encoded microRNAs (miRNAs) have been described in Bombyx mori nucleopolyhedrovirus; however, most of their functions remain unclear. Here we report the identification and characterization of an miRNA encoded by Autographa californica nucleopolyhedrovirus. The identified miRNA, AcMNPV-miR-1, perfectly matched a segment in the coding sequence of the viral gene ODV-E25 and downregulated ODV-E25 mRNA expression, which likely resulted in a reduction of infectious budded virions and accelerated the formation of occlusion-derived virions.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)