e23138 Background: While multi-cancer early detection (MCED)—a promising approach using one blood sample to detect multiple cancers—could supplement or replace standard-of-care (SOC) screening, its optimal position in population-based screening paradigm remains uncertain. This study aimed to evaluate the effectiveness and cost-effectiveness of integrating MCED at various phases of the screening process. Methods: We recalibrated the National Cancer Center (NCC) modeling framework to align with the latest population-based empirical data. The calibrated model was then used to simulate the natural history of the five highest-mortality cancers in China (lung, liver, colorectal, esophageal, and stomach) over a lifetime horizon for the entire Chinese population, with screening targeted at ages 50–74.The evaluated screening scenarios included no screening, risk-based SOC screening, and four screening strategies incorporating MCED: MCED used as a risk-assessment method (S1), MCED used as a clinical screening method (S2), MCED interception followed by SOC triage for MCED-negative individuals (S3), and MCED used as a supplemental screening for questionnaire non-high-risk individuals and SOC non-compliers (S4). The primary outcome was the 5% discounted incremental cost-effectiveness ratio (ICER), with a willingness-to-pay (WTP) threshold of three times per-capita GDP ($42,857 per quality-adjusted life-year [QALY] gained). Secondary outcomes included stage III-IV cancer diagnoses, cancer deaths, number needed to screen (NNS) to prevent one death. Sensitivity analyses were performed to assess the probability of cost-effective. Results: Compared with no screening, discounted ICERs for risk-based SOC and MCED strategies S1 to S4 were $1,808, $52,195, $26,726, $26,876, and $16,595 per QALY, respectively. Versus risk-based SOC, the ICERs for S4 and S3 were $33,809/QALY and $39,761/QALY. When using risk-based SOC as reference, S3 reduced stage III–IV cancer diagnoses by 6.9% and cancer mortality by 4.3%, while also improving screening efficiency by reducing the NNS from 240 to 81. S4 provided intermediate benefits with a 1.8% reduction in mortality, whereas S1 and S2 yielded smaller mortality reductions (<1.2%). Probabilistic sensitivity analysis indicated that S3 had the highest probability of being cost-effective at the WTP threshold (43.9%). Conclusions: Using MCED interception followed by SOC triage for MCED-negative individuals (S3) likely represents the most appropriate position for MCED in the cancer screening paradigm. In contrast, using MCED as a supplemental screening for questionnaire non-high-risk individuals and SOC non-compliers (S4) offers a lower-cost but suboptimal alternative.

HMGB1 (high mobility group protein) has been established as a key inflammatory mediator associated with various chronic diseases, particularly in neuroinflammatory pathogenesis. In this work, the potential of chrysin (Chr)/ apigenin (Api)/luteolin (Lut) as natural functional food ingredients was evaluated on their interactions with HMGB1 through integrated multi-spectroscopic analysis. Surface plasmon resonance (SPR) results of Api and Lut showed the dissociation constants (KD) values of 2.991 x 10-5 M and 3.206 x 10-6 M, respectively, which indicated that the hydroxyl group on the B-ring of flavonoids may impact their binding affinity with HMGB1. Fluorescence spectroscopy and circular dichroism (CD) analyses revealed that the binding of three flavonoids to HMGB1 caused static quenching and significantly altered their spatial conformation, resulting in a reduction in the alpha-helix content from 60.71 f 1.30% to Chr(44.24 f 1.27%), Api(47.52 f 2.79%) and Lut(49.20 f 2.07%), respectively. The synchronous and three-dimensional fluorescence spectrum further revealed that these interactions led to the microenvironmental changes in the chromogenic amino acids in HMGB1. Molecular docking experiments indicated that hydrogen bonding was the primary force between the interaction of three flavonoids and HMGB1, and the key amino acids forming the hydrogen bonds included Lys94, Lys95, Asp98, and Arg104 residues. Furthermore, the three flavonoids could mitigate HMGB1-induced neuroinflammation on BV2 microglia cells in the bioassay. Due to the key role of HMGB1 in neuroinflammation and neurological disorders, Chr, Api, and Lut could be applied to improve the treatment of neurological diseases, and they also provided new insights for the development of new dietary supplements of flavonoids.

3596 Background: Colorectal cancer (CRC) is the second most frequently diagnosed cancer in China and early detection could prevent over 90% of CRC-related deaths. Blood-based tests that analyze molecular features of CRC cell-free DNA (cfDNA), such as methylation and fragmentation patterns, hold great promise for early detection. However, the impact of molecular characteristics related to tumor location or mismatch repair (MMR) status on test performance has not been thoroughly investigated. In this study, we developed a blood-based CRC early detection test and analysed its performance across different CRC subgroups. Methods: A targeted enzymatic methyl sequencing panel was developed to identify tumor-specific hyper- and hypo-methylation markers and fragmentation profiles. A case-control cohort of 536 participants (268 CRC patients, 268 controls) was enrolled and startified into training and validation sets base on case/control status and cancer stage with 5-fold cross-validation. A gradient-boosted tree model was built by combining probabilities from methylomic and fragmentomic features. The optimal cutoff value for the early detection was determined by Youden's index, High specificity and High sensitivity methods, respectively. Results: The overall performance of Youden's index, High specificity and High sensitivity methods was as follows: specificity of 93.7%, 99.3%, 90.3%, and sensitivity of 96.6%, 86.2%, 97.0%, respectively. The area under the curve (AUC) value is 0.989 (95% CI: 0.981-0.996) , which is higher than those in current reports. When employing the High specificity method, the sensitivities were comparable between left and right-sided colon cancer (86.3% vs 85.7%, p = 1.0), and also similar between the dMMR (deficient mismatch repair) and pMMR (proficient mismatch repair) (87.5% vs 85.8%, p = 1.0), indicating that this model is applicable to various CRC subtypes. Additionally the TNM staging, pathological differentiation status, and the expression of Ki67, which are closely related to aggressiveness, were correlated with the sensitivity (Table). Conclusions: We have established CRC early detection model based on ctDNA methylation and fragmentation profiles, which shows excellent overall performance. Notably, this newly developed blood-based model shows no significant differences in sensitivity between distinct tumor locations or varying MMR statuses, suggesting its broader applicability across different types of CRC. Subgroup Positive/Total no. Sensitivity p_value Left-sidedRight-sided 195/22636/42 86.3%85.7% 1 dMMR pMMR 7/8200/233 87.5%85.8% 1 Stage IStage II Stage III Stage IV 34/5092/10872/7633/34 68%85.2%94.7%100% ＜0.001 Well differentiated Moderately differentiated Poorly differentiated 3/3160/19241/42 100%83.3%97.6% 0.029 Ki67_highKi67_low 138/15932/42 86.8%76.2% 0.147

4133 Background: In China, the 5-year survival rate of liver cancer patients is only 14%, far lower than the average of 43.7% for all cancer types. Early diagnosis and treatment are essential for survival. Traditional screening methods like AFP combined with abdominal ultrasound have low sensitivity. Recent studies suggest that blood cell-free DNA (cfDNA) characteristics could be a new screening approach for liver cancer. This study aims to compare methylation and fragmentation signals among liver cancer, hepatitis, cirrhosis patients, and healthy individuals, innovatively using these signals to construct an early-detection model which could improve patient prognosis. Methods: From July 2023 to November 2024, 315 blood samples were prospectively collected from five Chinese hospitals. The sample set included 105 liver cancer patients and 210 non-liver cancer controls (33 hepatitis, 30 cirrhosis, 147 healthy). This multi-center, multi-disease-controlled collection provides a robust data basis. Targeted enzymatic methyl sequencing detected over 600,000 methylation sites, enabling precise exploration of liver-cancer-related methylation. Beyond methylation, novel fragmentation features like break-point motifs, end motifs, arm-level count, fragment-size distribution and ratio were obtained. These, combined with methylation data, offer a multi-dimensional view for studying liver cancer pathogenesis and biomarkers. A gradient-boosted tree model, integrating 3840 methylation DMR features and fragmentomic model-predicted probabilities, was built. A nested cross-validation framework was used to optimize the model and ensure result accuracy. Results: The model achieved a high AUC of 0.97(95%CI:0.95-0.99) in liver cancer detection. At 96.2% specificity, the model had a 91.4% sensitivity for overall liver cancer detection, with 83.7% and 95.8% sensitivity for stage I and II respectively. Among 63 patients with hepatitis or cirrhosis, the model accurately predicted negative results in 88.9% of patients. Notably, for patients hard to identify by traditional tumor markers like AFP and DCP, the model showed high detection rates. When AFP < 400 ng/ml, the detection rate was 88.9%, and with concurrent DCP < 40 ng/ml, it reached 87.0%. When AFP < 20 ng/ml, the detection rate was 89.5%, and with DCP < 40 ng/ml simultaneously, the detection rate was 83.3%. Conclusions: This study established an early-detection model for liver cancer by leveraging cfDNA methylation and fragmentation signals. The model demonstrated remarkable performance, particularly in detecting liver cancer patients who are difficult to identify through conventional methods. It blazes a new trail for the early detection of liver cancer and could significantly enhance patient prognosis.

10536 Background: The global cancer burden is increasing, yet emerging risk factors for cancer incidence have not been comprehensively summarized in terms of the population attributable fraction (PAF) from a global perspective. Methods: We first searched and screened cancer risk factors from 435,977 studies in the Embase, PubMed, and Cochrane databases. Cancer risk effect sizes, such as RR (relative risk), HR (hazard ratio), and IRR (incidence rate ratio) values, were then obtained from meta-analyses. We further estimated population attributable fractions (PAFs) for each risk factor, cancer type, gender, SDI (Socio-demographic Index), modifiability, and trends from 1990 to 2021, using 94 pre-defined risk factors and 39 cancer types. These risk factors include lifestyle factors, environmental exposures, occupational risks, metabolic factors, comorbidities, and family or cancer history. Results: A total of 811 cancer risk effect sizes were obtained covering 75 countries or regions, including 18,084 cancer RR/HR/IRR values. In 2021, 64.1% (56.7% - 71.4%) of global cancer incidences were attributable to the evaluated risk factors, with modifiable risk factors accounting for 57.6%. Among the evaluated cancers, the proportion of incidence cases attributed to risk factors exceeded 50% in 26 types of cancer, with the leading cancers being cervical cancer (97.1%), Kaposi's sarcoma (91.2%), vaginal cancer (87.0%), lung cancer (84.6%), and laryngeal cancer (83.2%). In terms of individual risk factors, tobacco use (17.9%), infection (16.1%), and overweight or obesity (10.2%) contributed the highest PAFs, followed by family and genetic history (6.9%), alcohol consumption (5.7%), cardiovascular diseases (5.0%). The high SDI region shows a higher PAF (68.2% [61.5%–75.0%]) compared to the non-high SDI region (59.6% [52.2%–67.0%]). From 1990 to 2021, the global cancer incidences attributable to risk factors increased from 61.8% to 64.1%. Overweight or obesity, neurological and psychological disorders, and dietary factors were the top three leading increasing risk factors. Conclusions: The global cancer burden is significantly influenced by a wide range of modifiable and non-modifiable risk factors, with over 64% of cancer incidence in 2021 attributable to these factors. The rising trends in overweight or obesity, neurological and psychological disorders, and diabetes factors between 1990 and 2021 highlight the urgent need for targeted interventions to address these rapidly growing risks and reduction of the cancer burden.

Functional regulation of proteins by ligand-protein interactions plays a crucial role in understanding biological processes and identifying potential drugs. High mobility group box 1 (HMGB1) plays a pivotal role in sterile inflammation as a key immunomodulatory protein. Genistein, a well-known isoflavone compound, has been shown to have neuroprotective effects. In this study, we investigated the genistein-HMGB1 interactions using experimental and computational approaches. Our results revealed that genistein binds to HMGB1 with a KD value of 6.06 × 10-5 M. The addition of genistein significantly quenched the fluorescence of HMGB1. Thermodynamic analyses demonstrated that hydrogen bonds and hydrophobic forces are the primary forces during the binding process. Furthermore, the interaction between genistein and HMGB1 led to changes in the microenvironment of protein chromogenic amino acids and subtle alterations in the protein secondary structure. Molecular modeling results indicate that Pro95, Pro98, and Lys154 are the major amino acid residues for genistein binding to HMGB1. Meanwhile, at the cellular level, an inhibitory effect of genistein on HMGB1-induced NO release from microglia was observed, demonstrating an inhibition rate of 42.1 %. Our studies demonstrated that genistein could be applied in treating neurological diseases through its interaction with HMGB1.

Abstract Background  Spinal cord hemangioblastomas are rare benign and highly vascular tumors that develop either sporadically or as part of von Hippel–Lindau (VHL) disease. Generally, complete resection without significant neurologic deficit remains considerably challenging due to the risk of massive bleeding. The current study therefore aimed to describe en bloc resection of spinal cord hemangioblastomas according to the typical anatomical structures of peripheral lesions and evaluate the neurofunctional prognosis of this technique. Methods  A total of 39 spinal cord hemangioblastomas from a series of 19 patients who underwent en bloc resection were retrospectively analyzed. In all cases, clinical and radiologic characteristics, as well as surgical tenets, were retrospectively determined and analyzed. Short- and long-term outcomes were analyzed using the McCormick grade and Odom's criteria. Factors significantly associated with poor neurologic function after en bloc resection were also determined. Results  All 39 spinal cord hemangioblastomas, including 28 intramedullary, 2 intramedullary–extramedullary, and 9 extramedullary lesions, were located dorsally or dorsolaterally (100.0%). The most common lesion location was the thoracic segment (53.8%), with most of the lesions being accompanied by syringomyelia (94.7%). Long-term follow-up (mean: 103 ± 50.4 months) for prognosis determination revealed that 88.2% (15/17) of all cases had stable or improved neurofunctional outcomes according to the McCormick grade and Odom's criteria. Only one case with VHL disease developed recurrence 4 years after surgery. Additionally, statistical analysis showed that VHL disease was an independent prognostic factor associated with deteriorating neurologic function ( p  = 0.015). Conclusions  En bloc resection facilitated satisfactory long-term functional outcomes in patients with spinal cord hemangioblastomas. Given that VHL disease was identified as a predictor of poor long-term outcomes, regular long-term follow-up of patients with VHL-associated spinal cord hemangioblastoma seems necessary.

Introduction: Glioblastoma is a primary intracranial tumour with extremely high disability and fatality rates among adults. Existing diagnosis and treatment methods have not significantly improved the overall poor prognosis of patients. Nifuroxazide, an oral antibiotic, has been reported to act as a tumour suppressor in a variety of tumours and to participate in the process of antitumour immunity. However, whether it can inhibit the growth of glioma is still unclear. Methods: We explored the potential mechanism of nifuroxazide inhibiting the growth of glioblastoma cells through in vitro and in vivo experiments. Results: nifuroxazide can inhibit the proliferation of glioblastoma cells, promote G2 phase arrest, induce apoptosis, and inhibit epithelial-mesenchymal transition through the MAP3K1/JAK2/STAT3 pathway. Similarly, clinical sample analysis confirmed that MAP3K1 combined with STAT3 can affect the prognostic characteristics of patients with glioma. In addition, nifuroxazide can drive the M1 polarization of microglioma cells, inhibit the expression of CTLA4 and PD-L1 in tumour cells, and promote the infiltration of CD8 T cells to exert antitumour effects. Combination treatment with PD-L1 inhibitors can significantly prolong the survival time of mice. Conclusion: we found that nifuroxazide can inhibit the growth of glioblastoma and enhance antitumour immunity. Thus, nifuroxazide is an effective drug for the treatment of glioblastoma and has great potential for clinical application.

Data summary of methyl-sensitive cut counting sequencing (MSCC) sequencing of the 4 samples (8 libraries).

The natural product pectolinarigenin exerts anti-inflammatory activity and anti-tumor effects, and exhibits different biological functions, particularly in autophagy and cell cycle regulation. However, the antineoplastic effect of pectolinarigenin on glioblastoma (GBM) remains unclear. In the present study, we found that pectolinarigenin inhibits glioblastoma proliferation, increases autophagic flux, and induces cell cycle arrest by inhibiting ribonucleotide reductase subunit M2 (RRM2), which can be reversed by RRM2 overexpression plasmid. Additionally, pectolinarigenin promoted RRM2 protein degradation via autolysosome-dependent pathway by increasing autophagic flow. RRM2 knockdown promoted the degradation of CDK1 protein through autolysosome-dependent pathway by increasing autophagic flow, thereby inhibiting the proliferation of glioblastoma by inducing G2/M phase cell cycle arrest. Clinical data analysis revealed that RRM2 expression in glioma patients was inversely correlated with the overall survival. Collectively, pectolinarigenin promoted the degradation of CDK1 protein dependent on autolysosomal pathway through increasing autophagic flux by inhibiting RRM2, thereby inhibiting the proliferation of glioblastoma cells by inducing G2/M phase cell cycle arrest, and RRM2 may be a potential therapeutic target and a prognosis and predictive biomarker in GBM patients.

Genome instability is a hallmark of tumors and is involved in proliferation, invasion, migration, and treatment resistance of many tumors. However, the relationship of genome instability with gliomas remains unclear. Here, we constructed genome instability-derived long non-coding RNA (lncRNA)-based gene signatures (GILncSig) using genome instability-related lncRNAs derived from somatic mutations. Multiple platforms were used to confirm that the GILncSig were closely related to patient prognosis and clinical characteristics. We found that GILncSig, the glioma microenvironment, and glioma cell DNA methylation-based stemness index (mDNAsi) interacted with each other to form a complex regulatory network. In summary, this study confirmed that GILncSig was an independent prognostic indicator for patients, distinguished high-risk and low-risk groups, and affected immune-cell infiltration and tumor-cell stemness indicators (mDNAsi) in the tumor microenvironment, resulting in tumor heterogeneity and immunotherapy resistance. GILncSig are expected to provide new molecular targets for the clinical treatment of patients with gliomas.

The tumor microenvironment plays an important role in tumor progression. Hyaluronic acid (HA), an important component of the extracellular matrix in the tumor microenvironment, abnormally accumulates in a variety of tumors. However, the role of abnormal HA accumulation in glioma remains unclear. The present study indicated that HA, hyaluronic acid synthase 3 (HAS3), and a receptor of HA named CD44 were expressed at high levels in human glioma tissues and negatively correlated with the prognosis of patients with glioma. Silencing HAS3 expression or blocking CD44 inhibited glioma cell proliferation in vitro and in vivo. The underlying mechanism was attributed to the inhibition of autophagy flux and maintaining glioma cell cycle arrest in G1 phase. More importantly, 4-methylumbelliferone (4-MU), a small competitive inhibitor of Uridine diphosphate (UDP) with the ability to penetrate the blood-brain barrier (BBB), also inhibited glioma cell proliferation in vitro and in vivo. Thus, approaches that interfere with HA metabolism by altering the expression of HAS3 and CD44 and the administration of 4-MU potentially represent effective strategies for glioma treatment.

Purpose The identification of HER2 overexpression in a subset of gastric adenocarcinoma (GA) patients represents a significant step forward in unveiling the molecular complexity of this disease. The predictive and prognostic value of HER2 amplification in advanced HER2 inhibitor-treated GA patients has been investigated. However, its predictive value in resectable patients remains elusive. Methods We enrolled 98 treatment-naïve resectable Chinese GA patients with HER2 overexpression assessed using IHC. Capture-based targeted sequencing using a panel consisting of 41 gastrointestinal cancer-related genes was performed on tumor tissues. Furthermore, we also investigated the correlation between HER2 copy number (CN) and survival outcomes. Results Of the 98 HER2-overexpressed patients, 90 had HER2 CN amplification assessed using next-generation sequencing, achieving 92% concordance. The most commonly seen concurrent mutations were occurring in TP53 , EGFR and PIK3CA . We found HER2 CN as a continuous variable was an independent predictor associated with DFS ( p = 0.029). Our study revealed HER2 CN-high patients showed a trend of intestinal-type GA predominant ( p = 0.075) and older age ( p = 0.07). The median HER2 CN was 15.34, which was used to divide the cohort into CN-high and CN-low groups. Patients with high HER2 CN had a significantly shorter DFS than patients with low HER2 CN ( p = 0.002). Furthermore, HER2 CN as a categorical variable was also an independent predictor associated with DFS in patients. Conclusion We elucidated the mutation spectrum of HER2-positive resectable Chinese GA patients and the association between HER2 CN and DFS. Our work revealed HER2 CN as an independent risk factor predicted unfavorable prognosis in HER2-positive GA patients and allowed us to further stratify HER2-positive resectable GA patients for disease management.

Purpose: Brain gliomas are the most common primary malignant tumors of the central nervous system and one of the leading causes of death in patients with intracranial tumors. The lncRNA RPL34-AS1 is significantly upregulated in glioma tissues. However, the biological function of RPL34-AS1, especially in proliferation in glioma, remains unclear. Methods: The role of RPL34-AS1 in proliferation and angiogenesis in glioma cells was investigated using the LN229, U87, and U251 glioma cell lines. The levels of RPL34-AS1 were detected using real-time quantitative reverse transcription polymerase chain reaction. CCK-8 and colony formation assays were performed to determine the role of RPL34-AS1 in proliferation and survival, and its role in angiogenesis was assessed by an endothelial tube formation assay. Changes in protein levels were assessed by western blotting. Results: RPL34-AS1 was upregulated in glioma tissues and was correlated with tumor grade. RPL34-AS1 expression was also higher in glioma cells than in normal astrocytes. Knockdown of RPL34-AS1 blocked glioma cell proliferation by inhibiting angiogenesis. This effect occurred through decreased ERK/AKT signaling. Conclusions: This study suggests that RPL34-AS1 affects cell proliferation and angiogenesis in glioma and therefore may potentially serve as a valuable diagnostic and prognostic biomarker and therapeutic target in patients with glioma.

Background: Fast progression (FP), hyperprogressive disease (HPD), and early death (ED) are the newly reported cancer progression patterns in response to immune checkpoint inhibitor (ICI) treatment. This study aimed to investigate the clinical and genomic characteristics of FP, HPD, and ED following the ICI treatment of advanced non-small cell lung cancer (NSCLC). Methods: We retrospectively reviewed 117 patients with advanced NSCLC who were treated with ICIs from March 2017 to October 2019. FP was defined as (I) time to treatment failure (TTF) <1.5 months; and (II) >= 50% increase in the sum of the longest diameter (SLD) of target lesions. HPD was defined as (I) TTF <2 months; and (II) >= 50% change in tumor growth rate compared with before ICI initiation. ED was defined as overall survival (OS) <3 months. Tissue samples from 18 FP/HPD/ED patients and 5 partial response (PR) patients were subjected to genomic profiling. Genomic data from 693 tumor mutational burden- and histology-matched lung cancer samples were retrieved from an internal database as a control. Results: FP, HPD, and ED occurred in 7.21%, 9.38%, and 11.97% patients, respectively. The progression-free survival was comparable among the 3 groups. The median overall survival for FP, HPD, and ED were 3.19, 11.2, and 1.84 months, respectively. The genomic landscape revealed 1 EGFR amplification, 1 ALK fusion, 6 KRAS mutations, 1 ERBB2 amplification, 1 MET amplification, and 1 RET fusion among the 18 patients with FP/HPD/ED. Compared with the Control group, ED patients showed higher mutation frequencies for KRAS (P<0.01), CDKN1B (P<0.01), and NTRK1 (P=0.04). Mutations in RAD54L (P=0.018) and MYC (P=0.04) were more common in FP patients; HPD patients showed more frequent RAD54L mutations (P<0.001). Conclusions: We demonstrated different genomic characteristics across different progression patterns following ICI treatment, which might assist clinicians in the prediction of a patient's response, identifying candidates for more effective ICI therapy.

Satisfactory tumor material is often hard to obtain for molecular analysis in extranodal natural killer (NK)/T-cell lymphoma (NKTCL) at present. However, the accuracy and utility of circulating cell-free DNA (cfDNA) genotyping have not been adequately assessed in NKTCL. We therefore performed targeted next-generation sequencing on tumor tissues and a series of longitudinal plasma samples prospectively collected from a cohort of high-risk NKTCL patients. Concordance of genotyping results of paired baseline tumor and cfDNA and the predictive value of dynamic cfDNA monitoring were evaluated. At baseline, 59 somatic variants in 31 genes were identified in tumor and/or plasma cfDNA among 19 out of 24 high-risk NKTCL patients (79.2%). Plasma cfDNA had a sensitivity of 72.4% for detection of somatic variants identified in tumor biopsies before treatment. Plasma cfDNA also allowed the identification of mutations that were undetectable in tumor biopsies. These results were also verified in a validation cohort of an additional 23 high-risk NKTCL patients. Furthermore, longitudinal analysis showed that patients with rapid clearance of NKTCL-related mutations from plasma had higher complete remission rates (80.0% vs 0%; P = .004) and more favorable survival (1-year progression-free survival [PFS] rate, 79.0% vs 20.0%; P = .002) compared with those with persisting or emerging mutations in plasma. In addition, low cfDNA concentration before treatment was associated with favorable survival outcome for patients with NKTCL (1-year PFS, 90.0% vs 36.4%; P = .012). In conclusion, cfDNA mirrors tumor biopsy for detection of genetic alterations in NKTCL and noninvasive dynamic plasma cfDNA monitoring might be a promising approach for tracking response and survival outcome for patients with NKTCL.

BACKGROUND:HER2 transmembrane domain (TMD) mutation has been reported as a rare driver mutation associated with advanced stage disease and a poor prognosis in patients with lung adenocarcinoma (LUAD). We aimed to comprehensively profile the genetic landscape and treatment response information of HER2 TMD-mutant LUAD.METHODS:An in-house database of 7,812 LUAD patients was screened for mutation prevalence. A multi-center cohort of 16 HER2 V659E-mutant patients and an external cohort of 38 HER2-mutant patients from cBioPortal with overall survival (OS) data were analyzed. Eight patients from the in-house cohort were included in the real-world study of treatment response. Molecular docking simulation and binding affinity prediction were performed.RESULTS:In Chinese LUAD, the prevalence of HER2 TMD mutation was 0.18% (14/7,812), and 0.14% (11/7,812) for the HER2 V659E mutation. The most recurrent co-alteration was TP53 mutation (n=4, 25%) and HER2 amplification (n=2, 12.5%). TMD-mutant patients were diagnosed at more advance stages (P<0.001) and had poorer OS (median OS 10.0 vs. 61.6 months, HR =7.9, 95% CI: 1.0-61.0, P<0.001) than non-TMD mutations. The overall response rate of targeted therapy, chemo-based therapy, and immunotherapy was 57.1%, 22.2%, and 0%, respectively. We postulated to challenge the resistance of tyrosine kinase inhibitor (TKI) with another with stronger binding energy to HER2 and supported the conclusion with a successful case. Additionally, we demonstrated a three-month response to the off-label use of pyrotinib in fifth-line therapy.CONCLUSIONS:Comapred with non-TMD mtuations, HER2 TMD mutation is a rare driver mutation with poorer prognosis in LUAD. Targeted therapy is the dominant choice for patients harboring this targetable mutation and longer OS could possibly be achieved through rechallenge with TKI of stronger binding affinity. Response to fifth-line pyrotinib was observed.

Background: The genomic profile of non-small cell lung cancer (NSCLC) in Asians is distinct from that of Caucasians, but comprehensive genetic profiling reports have been limited for Asian patients. We aimed to elucidate genomic characteristics of Chinese NSCLC patients and develop potential model including genomic characteristics to predict postoperative prognosis. Methods: Resected tumor samples from 511 patients with stage I-IV lung cancer were subjected to targeted sequencing using a panel of 295 cancer-related genes. Based on the molecular profiles and clinical features, we established nomogram models with predictors consisting of integrated clinical and genomic characteristics to provide post-operative risk stratification. Results: Compared to the TCGA population (mainly Caucasians), there was a significantly higher frequency of EGFR (53.7% vs. 14.4%) and NOTCH3 (8.4% vs. 1.3%) mutations and less mutated KRAS (11.0% vs. 32.6%), KEAP1 (4.4% vs. 17.4%) and LRP1B (16.3% vs. 29.6%) in Chinese lung adenocarcinomas (LUAD). Distinct patterns of mutually exclusive and co-occurring mutations were identified between LUAD and lung squamous cell carcinoma (LUSC), indicating the unique histology-specific tumorigenesis mechanism of each subtype. We observed alterations in pathways correlated with clinical characteristics. Additionally, we constructed nomogram model with predictors consisting of clinical and genomic characteristics, which were more accurate than models with clinical characteristics or TNM staging only both in stage I-IIIA patients and T1-2N0M0 sub-cohort. Conclusions: This study revealed Chinese NSCLC patients have unique genomic profile. Furthermore, the nomogram model combining clinical features with genomic characteristics could improve risk stratification in early-stage NSCLC.

INTRODUCTION:The optimal treatment for EGFR-mutant lung adenocarcinoma (LUAD) remains challenging because of intratumor heterogeneity. We aimed to explore a refined stratification model based on the integrated analysis of circulating tumor DNA (ctDNA) tracking. METHODS:ctDNA was prospectively collected at baseline and at every 8 weeks in patients with advanced treatment-naive EGFR-mutant LUAD under gefitinib treatment enrolled in a phase 2 trial and analyzed using next-generation sequencing of a 168-gene panel. RESULTS:Three subgroups categorized by baseline comutations-EGFR-sensitizing mutations (59, 32.8%), EGFR-sensitizing mutations with tumor suppressor mutations (97, 53.9%), and EGFR-sensitizing mutations with other driver mutations (24, 13.3%)-exhibited distinct progression-free survival (13.2 [11.3-15.2] versus 9.3 [7.6-10.5] versus 4.0 [2.4-9.3] months) and overall survival (32.0 [29.2-41.5] versus 21.7 [19.3-27.0] versus 15.5 [10.5-33.7] months, respectively), providing evidence for initial stratification. A total of 63.7% of the patients achieved week 8 ctDNA clearance, with significant difference noted among the three subgroups (74.5% versus 64.0% versus 29.4%, respectively, p = 0.004, Fisher's exact test). Patients without week 8 ctDNA clearance had worse progression-free survival (clearance versus nonclearance 11.2 [9.9-13.2] versus 7.4 [5.6-9.6] months, p = 0.016, Cox regression], especially in the second subgroup [5.8 (5.6-11.5) months], suggesting the necessity of adaptive stratification during treatment. During follow-up, 56.0% and 20.8% of the patients eventually harbored p.T790M and non-p.T790M mutations, respectively, with a significant difference in non-p.T790M mutations among the three subgroups (7.5% versus 15.7% versus 80.0%, respectively, p < 0.001, Fisher's exact test), giving clues to postline treatment. CONCLUSIONS:The patients with baseline comutations and ctDNA nonclearance at first visit might require combined therapy because of the limited survival benefit of EGFR tyrosine kinase inhibitor monotherapy. We proposed a refined stratification mode for the whole-course management of EGFR-mutant LUAD.

Objectives: Although the majority of epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC) patients respond to EGFR tyrosine kinase inhibitors (TKIs), significant heterogeneity in clinical response is observed which might be attributed to the distinct sub-molecular characteristics. The present study aims to identify genetic alterations correlated with clinical outcomes and treatment response to different EGFR-TKI inhibitors. Materials and methods: We integrated the genomic data and clinical outcomes including progression-free survival (PFS) and overall survival (OS) in 179 patients with advanced EGFR-mutant NSCLC who were treated with EGFR-TKI as 1st line of treatment. Results: We found that EGFR-mutant patients harboring concomitant TP53 mutation (OS: 21 vs. 40 months, P = 0.05), ERBB2 amplification (PFS: 6.1 vs. 12.5 months, P = 0.01) or FGF19 amplification (OS: 11.2 vs. 27.1 months, P = 0.01) were significantly associated with a poorer clinical prognosis after treated with 1st generation EGFR-TKI. In contrast, the presence of TP53 mutation did not affect the PFS nor OS of patients treated with 2nd generation EGFR-TKI. Furthermore, EGFR-mutant and TP53-wild type (WT) patients benefited more from a combinatorial treatment consisting of EGFR-TKI and bevacizumab comparing to EGFR-TKI as a single agent (PFS: 21.7 vs. 9.3 months, P < 0.01). Copy number variation (CNV) (PFS: 4.6 vs.9.4 months, p = 0.018) was identified as an unfavorable predictive factor to 3rd-generation TKI. We also revealed distinct resistance mechanisms associated with different EGFR-TKIs. Conclusion: Our study highlights the heterogeneity both in the primary molecular landscape and acquired alterations in EGFR-mutated NSCLCs, which might play a role in determining the clinical efficacy of EGFR-TKIs. We also revealed the differential prognostic role of TP53 mutation in patients treated with the 1st or 2nd generation of EGFR-TKI. Our study also suggests that EGFR-mutant and TP53-WT patients may benefit more from combinatorial treatment consisting of EGFR-TKI and bevacizumab, highlighting the importance of further stratifying EGFR-mutant patients.