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BACKGROUND: Comprehensive genomic profiling has become standard clinical practice in the management of advanced lung cancer. In addition to tissue and plasma, other body fluids are also being actively explored as alternative sources of tumor DNA. This study investigated the utility of induced sputum obtained from patients with non-small-cell lung cancer (NSCLC) for somatic variation profiling. METHODS: Our study included 41 treatment-naïve patients diagnosed with locally advanced to advanced NSCLC between October 2018 and June 2019. Capture-based targeted sequencing was performed on matched tumor, plasma, and induced sputum samples of 41 patients using a 168-gene panel. We analyzed the somatic variations detected from each sample type and the concordance of variations detected between matched samples. The concordance rate was defined as the proportion of the total number of variations detected from one sample type relative to the reference sample type. RESULTS: Comparative analysis on the somatic variation detection using matched tumor samples as a reference revealed detection rates of 76.9% for plasma, 72.4% for sputum-supernatant, and 65.7% for sputum-sediment samples. Plasma, sputum-supernatant, and sputum-sediment achieved positive predictive values of 73.3%, 80.4%, and 55.6% and sensitivities of 50.0%, 36.9%, 31.3%, respectively, relative to tumor samples for 168 genes. Sputum-supernatants had significantly higher concordance rates relative to matched tumor samples (69.2% vs. 37.8%; P=0.031) and maximum allelic fraction (P

BackgroundPulmonary neuroendocrine tumors (pNETs) include typical carcinoid (TC), atypical carcinoid (AC), large cell neuroendocrine carcinoma (LCNEC), and small cell lung carcinoma (SCLC). The optimal treatment strategy for each subtype remains elusive, partly due to the lack of comprehensive understanding of their molecular features. We aimed to explore differential genomic signatures in pNET subtypes and identify potential prognostic and therapeutic biomarkers.MethodsWe investigated genomic profiles of 57 LCNECs, 49 SCLCs, 18 TCs, and 24 ACs by sequencing tumor tissues with a 520-gene panel and explored the associations between genomic features and prognosis.ResultsBoth LCNEC and SCLC displayed higher mutation rates for TP53, PRKDC, SPTA1, NOTCH1, NOTCH2, and PTPRD than TC and AC. Small cell lung carcinoma harbored more frequent co-alterations in TP53-RB1, alterations in PIK3CA and SOX2, and mutations in HIF-1, VEGF and Notch pathways. Large cell neuroendocrine carcinoma (12.7 mutations/Mb) and SCLC (11.9 mutations/Mb) showed higher tumor mutational burdens than TC (2.4 mutations/Mb) and AC (7.1 mutations/Mb). 26.3% of LCNECs and 20.8% of ACs harbored alterations in classical non-small cell lung cancer driver genes. The presence of alterations in the homologous recombination pathway predicted longer progression-free survival in advanced LCNEC patients with systemic therapy (P = .005) and longer overall survival (OS) in SCLC patients with resection (P = .011). The presence of alterations in VEGF (P = .048) and estrogen (P = .018) signaling pathways both correlated with better OS in patients with resected SCLC.ConclusionWe performed a comprehensive genomic investigation on 4 pNET subtypes in the Chinese population. Our data revealed distinctive genomic signatures in subtypes and provided new insights into the prognostic and therapeutic stratification of pNETs.

Diagnosing multiple lung tumors as multiple primary lung cancer (MPLC) or intrapulmonary metastasis (IPM) has important therapeutic implications. The clinical applicability of next-generation sequencing (NGS) has been explored, although there is currently no consensus regarding how to integrate its findings for cancer staging. We performed a retrospective analysis of the mutational landscape of 13 patients with multiple tumors and underwent surgery at Renmin Hospital of Wuhan University from Sep. 2018 to Dec. 2020.The cohort consisted mostly of women (11/13, 84.6%) and non-smokers (12/13, 92.3%). Median age was 58 years (range 26-71). No patient manifested relapse after a median follow-up duration of 11.2 months (range 8.2-36.0). NGS was performed with a large panel (≥ 350 genes) in most cases (11/13, 84.6%). Most nodules were invasive adenocarcinomas (25/41, 61.0%), followed by adenocarcinomas in situ (13/41, 31.7%), and minimally invasive adenocarcinomas (3/41, 7.3%). Four patients were clinically diagnosed with IPM, of whom 1 had only one nodule sequenced. Of the 12 patients with genomic profiles from multiple nodules, 10 (83.3%) had consistent NGS inference and clinical diagnosis of MPLC or IPM. Of the remaining two, P1 was clinically diagnosed with IPM but NGS did not detect any genomic aberrations from one nodule, which may have missed evidence that could have led to inference of clonal relationship. P8 was diagnosed with MPLC but had two nodules that harbored EGFR L858R mutation. NGS revealed clonal relationships that enabled high-confidence differential diagnosis in the concordant cases. For instance, the 3 nodules in P10 each harbored an oncogenic driver: EGFR exon 19 deletion, EGFR L858R, and ALK fusion, which were suggestive of tumor clones of separate origins. Similar mutual exclusion was also observed in P6 (EGFR exon 19 deletion vs L858R), P2 (EGFR exon 19 deletion vs MAP2K1), and P7 (2 EGFR exon 18 mutations G719A and E709 in one nodule vs EGFR E746_A750del in another). On the other hand, P9 illustrates a case of complex clonal architecture, in which 8 nodules were derived from 5 origins. Further analysis found that alterations in KRAS were associated with smaller nodule size, whereas those in TP53, CDKN2A, NKX2-1, or GNAS were associated with greater size.In summary, NGS achieved highly concordant MPLC/IPM differentiation with clinical diagnosis based on histopathologic and radiographic evidence. However, caution is warranted when NGS detects no aberration from any of the multiple sequenced nodules. Examination of the mutational landscape found that altered proto-oncogene KRAS was associated with smaller tumors whereas altered tumor suppressors TP53 and CDKN2A associated with smaller ones, suggesting different roles in these molecules play in tumor growth. Citation Format: Wenyang Jiang, Wei Wang, Luting Qiu, Xiao Zou, Zhiqiu Chen, Qing Geng. Next-generation sequencing revealed clonal architecture and faciliated diagnosis of patients with multiple lung tumors: An analysis of 12 cases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5232.

Background Comprehensive mutation profiling has become a standard clinical practice in the management of advanced lung cancer. In addition to tissue and plasma, other body fluids are also being actively explored as alternative sources of tumor DNA. In this study, we investigated the potential of induced sputum obtained from patients with non-small cell lung cancer (NSCLC) for mutation profiling. Methods Capture-based targeted sequencing was performed on matched tumor, plasma, and induced sputum samples of 41 treatment-naïve patients with NSCLC using 168 gene panel. Results Comparative analysis on the mutation detection using matched tumor sample as reference revealed detection rates of 76.9% for plasma, 72.4% for sputum-supernatant, and 65.7% for sputum-sediment samples. Plasma, sputum-supernatant, and sputum-sediment achieved positive predictive values of 73.3%, 80.4%, and 55.6% and sensitivities of 50.0%, 36.9%, 31.3%, respectively, relative to tumor samples for 168 genes. Sputum-supernatants had significantly higher concordance rates relative to matched tumor samples (69.2% vs 37.8%; P = 0.031) and maximum allelic fraction (P P = 1) but with significantly higher maximum allelic fraction than their matched plasma samples (P = 0.003). Furthermore, sputum-supernatant from smokers had a significantly higher maximum allelic fraction than sputum-supernatant from non-smokers (P = 0.021). Conclusions Our study demonstrated that supernatant fraction from induced sputum is a better sampling source than its sediment and has comparable performance as plasma samples. Induced sputum from NSCLC patients could serve as an alternative media for next-generation sequencing-based mutation profiling.

Radiation is known to be associated with pathology of various human diseases. This study has focused on the effect of radiation on glucose homeostasis with regard to metabolic function of liver and pancreas and the effect of polysaccharide from the fruiting bodies of Auricularia auricular (SNAAP) on glucose metabolism. The

Introduction It has been well established that EGFR Thr790Met is one of the major resistance mechanisms to first- and second-generation EGFR tyrosine kinase inhibitors (TKIs). Nevertheless, whether EGFR Thr790Leu (T790L), which shares the mutation site of Thr790 with EGFR Thr790Met, mediates resistance to EGFR TKIs remains elusive. The treatment options for patients harboring this rare mutation have not been reported. Methods Capture-based targeted ultradeep sequencing was performed on tumor and plasma samples collected at various treatment milestones from three patients with advanced lung adenocarcinoma undergoing targeted therapy. Results Needle biopsy of lymph node metastasis from patient 1 revealed EGFR T790L at disease progression on first-line treatment of gefitinib. Patient 2 had EGFR T790L identified from needle biopsy of lung tissue at disease progression on icotinib treatment. This patient was subsequently treated with osimertinib and achieved stable disease with a progression-free survival of 9 months. For patient 3, at disease recurrence after surgery, resected lung tumor tissue was retrieved for molecular profiling and revealed EGFR exon 19 deletion and EGFR T790L. The patient subsequently received osimertinib treatment and continued to benefit for 16 months and counting. She has maintained stable disease at the time of submission of this manuscript. Conclusions We revealed for the first time that EGFR T790L may serve as a potential resistance mechanism to first-generation EGFR TKIs. We also report the first clinical evidence of efficacy generated by osimertinib in patients with lung adenocarcinoma harboring primary or acquired EGFR T790L, shedding light on treatment options for this subset of patients.

9043 Background: MET amplification is an important mechanism mediating acquired resistance to EGFR tyrosine kinase inhibitors (TKI). Until now, no consensus exists on the standard treatment strategy for this subset of patients due to the lack of clinical data from large cohort or controlled trials. In our clinical practice, three regimens were commonly administered to patients after MET amplification-mediated EGFR-TKI progression: EGFR-TKI and MET-TKI combination therapy, MET-TKI monotherapy, or chemotherapy. Our study aimed to compare the effectiveness of these three regimens. Methods: Seventy patients with EGFR-mutant advanced NSCLC who progressed from prior EGFR-TKI through the acquisition of MET amplification and received treatment between March 2015 and March 2020 were included in this study. Of them, 38 received EGFR-TKI plus crizotinib, 10 received crizotinib monotherapy, and 22 received platinum-based doublet chemotherapy. Somatic mutation profiling was performed on blood and tissue biopsy samples. Resistance mechanisms to the combination targeted therapy were also explored in 12 patients. Results: The objective response rate (ORR) and disease control rate (DCR) were 47.5% and 84.0% for EGFR-TKI+crizotinib group, 40.0% and 70.0% for crizotinib monotherapy group, and 18.2% and 50.0% for chemotherapy group, respectively. The EGFR-TKI+crizotinib group had significantly better ORR (P = 0.026) and DCR (P = 0.016) than the chemotherapy group but was not statistically different from the crizotinib monotherapy group (ORR, P = 0.73; DCR, P = 0.39). Progression-free survival (PFS) was significantly longer for the EGFR-TKI+crizotinib group than those who received crizotinib monotherapy (5.0 vs 2.3 months, P = 0.004) or chemotherapy (5.0 vs 2.9 months, P = 0.036), but overall survival was comparable (10.0 vs 4.1 vs 8.5 months, P = 0.088). TP53 mutation (58.5%) and EGFR amplifications (42.9%) were the two common concurrent mutations in the three cohorts. PFS was significantly longer for patients with either concurrent TP53 mutation (n = 17) (6.0 vs 2.3 vs 2.9 months, P = 0.009) or concurrent EGFR amplification (n = 13) (5.0 vs 1.2 vs 2.4 months, P = 0.016) who received EGFR-TKI+crizotinib. Potential molecular mechanisms of acquired resistance to EGFR-TKI+crizotinib therapy included EGFR T790M (n = 2), EGFR L718Q (n = 1), EGFR S645C (n = 1), MET D1228H (n = 1), BRAF V600E (n = 1), NRAS Q61H (n = 1), and amplifications in KRAS (n = 2), ERBB2 (n = 1), CDK4 (n = 1), and MYC (n = 2). Conclusions: Our study provides real-world clinical evidence, in the largest cohort to date, that simultaneous inhibition of EGFR and MET improves clinical outcomes of patients with EGFR-mutant NSCLC who acquired MET amplification from prior EGFR-TKI therapy, indicating that combinatorial regimen of EGFR-TKI and MET-TKI could be a more effective therapeutic strategy in this subset of patients.

e21617 Background: With the advancements in the development of targeted therapy, the detection of actionable genes has become routine practice in diagnosing lung cancer. Plasma-based mutation profiling is widely used in clinical settings. Profiling using other body fluids is actively explored. In this study, we investigated the potential of induced sputum obtained from non-small cell lung cancer (NSCLC) patients for mutation profiling. Methods: We performed targeted sequencing on matched FFPE, plasma and induced sputum samples of 41 treatment-naïve advanced NSCLC using a panel consisting of 168 lung cancer-related genes. 10ml of whole blood and 8 ml of induced sputum were obtained from each patient. Induced sputum samples were fractionated into supernatant (sup) and precipitates (ppt). The cohort had a median age of 65 with a majority (32/41) diagnosed with lung adenocarcinoma. 7 of them were diagnosed with squamous cell carcinoma and the remaining 2 patients were diagnosed with neuroendocrine tumor. All patients had advanced disease (stage III or IV). Results: First, we investigated the optimal sputum fraction for mutation profiling by comparing the detection rates and maximum allelic fraction (maxAF) between sputum sup and ppt samples using matched tissue and plasma as references. The detection rate was defined as any mutation detected from the panel used; while maxAF was defined as the highest mutation allelic fraction among all mutations detected. Tissue, plasma, sputum sup and ppt achieved a rate of detection of 100%, 76.9%, 72.4% and 65.7% respectively, revealing comparable detection rates between plasma and sputum sup (p = 0.89). Sup had a significantly higher average maxAF than plasma (p = 0.03) and ppt samples (p < 0.01). Using driver mutations detected from tissue samples as references, sup, ppt and plasma samples achieved a sensitivity of 63.2%, 50% and 67.8%, respectively. Next, we investigated whether induced sputum obtained from smokers would be more informative than non-smokers. Our analysis revealed that sup from smokers had a significantly higher average AF than sup from non-smokers (p < 0.001). Furthermore, there was a statistical significant difference in average AF between sup and ppt obtained from smoker (p < 0.001) but such difference was non-existent in non-smokers (p = 0.46). Conclusions: Our study demonstrated that sputum sup demonstrates a comparable performance as plasma samples and is as a better sampling source than its ppt. Induced sputum from advanced stage NSCLC patients is an alternative media for mutation profiling.

The RAD18 E3 ligase requires SIVA1 as an accessory protein for binding to its substrate PCNA during translesion DNA synthesis., Translesion DNA synthesis (TLS) is a universal DNA damage tolerance mechanism conserved from yeast to mammals. A key event in the regulation of TLS is the monoubiquitination of proliferating cell nuclear antigen (PCNA). Extensive evidence indicates that the RAD6–RAD18 ubiquitin-conjugating/ligase complex specifically monoubiquitinates PCNA and regulates TLS repair. However, the mechanism by which the RAD6–RAD18 complex is targeted to PCNA has remained elusive. In this study, we used an affinity purification approach to isolate the PCNA-containing complex and have identified SIVA1 as a critical regulator of PCNA monoubiquitination. We show that SIVA1 constitutively interacts with PCNA via a highly conserved PCNA-interacting peptide motif. Knockdown of SIVA1 compromised RAD18-dependent PCNA monoubiquitination and Polη focus formation, leading to elevated ultraviolet sensitivity and mutation. Furthermore, we demonstrate that SIVA1 interacts with RAD18 and serves as a molecular bridge between RAD18 and PCNA, thus targeting the E3 ligase activity of RAD18 onto PCNA. Collectively, our results provide evidence that the RAD18 E3 ligase requires an accessory protein for binding to its substrate PCNA.

The diastereoselective reaction of aldimines (I) with silane (II) at low temperature give α-(dichloromethyl)amines (III) whereas aziridines (IV) are obtained when the reaction mixture is warmed up to room temperature.

From increasing the quality and reliability of military products, the new ensuring quality system for UAVs is established. Firstly, the current quality management for UAVs is analyzed. The focus and limitation of the total life-span quality management is introduced from developing, producing and equipping. Secondly, the quality and reliability information system is introduced on information searching, disposing and using. The design, key questions and mathematics models for statistic and forecast on Management Information System (MIS) are elaborated. Lastly, the quality management method of the total life-span condition and the reliability information management is introduced. The design and using of the system break traditional independence scheme on systems or subsystems, producers or users. The information collection extends to basic accessory, goes through the producers or users and feedbacks to after service quality control and new products developing. When the system used all information resources are integrate. The working efficiency on military UAVs maintenance is increased and the expense on total life-span maintenance is decreased.

According to the design requirement of UAV (the unmanned aircraft vehicle), the models of threat, UAV's dynamics, guidance and trajectory planning are established. Trajectory planning adopts genetic algorithms (GAs) and genetic coding uses "distance, corner" program. Through simulating by Matlab/Simlink software, the result proves the trajectory planning according with dynamics, characteristic.

The primary goal of this research is to develop a novel intelligent fault diagnosis method employing neuro-fuzzy transductive inference algorithm (NFTI) in order to solve the the global model application problem, as well as the global availability of the model and sample data set. The method is characterized by that a personal local model which is established for every new fault symptom input data in the fault diagnosis systems, based on some closest samples next to this fault symptom data in an existing sample database. Compared with other similar inductive method (ANFIS - adaptive neuro-fuzzy inference system) on Fisherpsilas Iris data set, the mentioned algorithm classifier has reduced 15% of the average test error and increased approximately 30% of classification speed. Detecting the fault symptom data set sampled from actual aeronautic thrustor test, the presented system can identify accurately three fault states. The results of the research indicate that the availability and efficacy of the fault diagnostic strategy is superior to any other inductive reasoning technique about some fault diagnosis issues.

The asymmetric unit of the title compound, C6H5Cl2NO2S, contains two molecules with similar conformations (r.m.s. overlay fit for the non-H atoms = 0.067 Å). Atoms attached to the pendent Csp3—S bond are arranged in a staggered conformation with one of the Cl atomsantito the C atom in the aromatic ring [C—S—C—Cl torsion angles = 178.41 (11) and −176.70 (13)°]. In the crystal, molecules are linked by C—H...N and C—H...O hydrogen bonds, generating a three-dimensional network, and weak aromatic π–π stacking is also observed [centroid–centroid separation = 3.8902 (17) Å].

Protocols for the stereoselective synthesis of α-(dichloromethyl)amines, α-(chloromethyl)amines, and α-chloroaziridines are presented. Diastereoselective synthesis of α-(dichloromethyl)amines was achieved based on nucleophilic dichloromethylation of aromatic N-tert-butylsulfinyl aldimines with (dichloromethyl)trimethylsilane at a low reaction temperature. Slowly warming the reaction mixture up to room temperature gave α-chloro cis-aziridines. Additionally, with Bu3SnH as the reductant, α-(dichloromethyl)amines were readily obtained from easily accessible α-(trichloromethyl)amines via mono-dechlorination. Over-reduction was successfully suppressed. Subsequent radical mono-dechlorination of the α-(dichloromethyl)amines gave the corresponding α-(chloromethyl)amines in good to excellent yields.