The identification of HER2-low and HER2-ultralow breast cancer (BC) subgroups has garnered considerable attention following the demonstrated clinical efficacy of HER2-targeted antibody-drug conjugates (ADCs). This study investigated whether these subgroups represent distinct biological subtypes compared to HER2-undetected tumors. Analyzing 297 HER2-negative BCs stratified by immunohistochemistry (50.2 % HER2-low, 35.7 % HER2-ultralow, 14.1 % HER2-undetected), we systematically compared clinicopathological and molecular profiles (AmoyDx® HANDLE Classic Panel). HER2-detected tumors (combined low/ultralow cohort) showed significant differences in hormone receptor (HR) positivity versus HER2-undetected tumors (p = 0.001). The mutation frequencies of the three most frequently altered genes (TP53, PIK3CA, and PTEN) in HER2-ultralow tumors did not significantly differ from those in the two other subgroups. However, HER2-detected tumors exhibited different molecular alterations compared to HER2-undetected tumors, with increased PIK3CA mutations in the former (p = 0.024) and TP53 mutation enrichment in the latter (p = 0.007). These differences attenuated when tumors were stratified by HR status. In addition, HER2 expression positively correlated with ERBB2 copy number, showing higher mean values in HER2-detected subgroups versus undetected counterparts (2.01 ± 0.31 vs 1.88 ± 0.23; p = 0.011). While no significant survival differences emerged across subgroups, most mortality events occurred in HER2-low (6/12, 50.0 %) and HER2-ultralow (5/12, 41.7 %) cases. Our findings indicate that while HER2-low and HER2-ultralow breast cancers may not represent independent biological subtypes, their clinical distinction from HER2-undetected tumors remains crucial due to potential ADC therapeutic implications.

The detection of anomalous tissue regions (ATRs) within affected tissues is crucial in clinical diagnosis and pathological studies. Conventional automated ATR detection methods, primarily based on histology images alone, falter in cases where ATRs and normal tissues have subtle visual differences. The recent spatial transcriptomics (ST) technology profiles gene expressions across tissue regions, offering a molecular perspective for detecting ATRs. However, there is a dearth of ATR detection methods that effectively harness complementary information from both histology images and ST. To address this gap, we propose MEATRD, a novel ATR detection method that integrates histology image and ST data. MEATRD is trained to reconstruct image patches and gene expression profiles of normal tissue spots (inliers) from their multimodal embeddings, followed by learning a one-class classification AD model based on latent multimodal reconstruction errors. This strategy harmonizes the strengths of reconstruction-based and one-class classification approaches. At the heart of MEATRD is an innovative masked graph dual-attention transformer (MGDAT) network, which not only facilitates cross-modality and cross-node information sharing but also addresses the model over-generalization issue commonly seen in reconstruction-based AD methods. Additionally, we demonstrate that modality-specific, task-relevant information is collated and condensed in multimodal bottleneck encoding generated in MGDAT, marking the first theoretical analysis of the informational properties of multimodal bottleneck encoding. Extensive evaluations across eight real ST datasets reveal MEATRD's superior performance in ATR detection, surpassing various state-of-the-art AD methods. Remarkably, MEATRD also proves adept at discerning ATRs that only show slight visual deviations from normal tissues.

Supplementary Table 8. The response rate by MYD88/CXCR4 genotype during first-line cytotoxic therapy.

Supplementary Figure 9. The survival of patients according to IPSSWM risk model. (A) The overall survival of the whole cohort; (B) The overall survival in patients treated with BTKi-based therapy

Supplementary Figure 2. The IGHV-D-J repertoire in patients with mutated and wild-type MYD88. Distribution of major VH family gene segments in wild-type group (A) and mutated group (B); Distribution of major DH family gene segments in wild-type group (C) and mutated group (D); Distribution of major JH family gene segments in wild-type group (E) and mutated group (F)

Supplementary Figure 8. The survival outcomes of patients with MYD88/CXCR4 mutation status according to the option of different first-line therapies. The progression free survival and overall survival in patients with cytotoxic therapy (A-B); in patients with Rtuximab/bortezomib-based therapy; (C-D); and in patients with BTKi-based therapy (E-F). *P < 0.05, **P < 0.01, ***P < 0.001.

Purpose/Objective(s) The prognosis of nasopharyngeal carcinoma with T4 or N3 remains unsatisfactory. The aim of this study was to assess the efficacy and safety of neoadjuvant with GP plus tislelizumab followed by concurrent chemoradiotherapy, and adjuvant treatment with tislelizumab, an anti-PD-1 monoclonal antibody, in previously untreated stage IVA (T4 or N3) nasopharyngeal carcinoma. Materials/Methods In this phase II, single-arm study, eligible patients aged 18-70 years who were diagnosed with stage IVA (AJCC 8th) non-keratinizing nasopharyngeal carcinoma received neoadjuvant therapy with gemcitabine (1000mg/m2 on day 1,8), cisplatin (25 mg/m2 on day 1-3) and tislelizumab (200mg) Q3W for 2 cycles followed by concurrent IMRT and cisplatin (100 mg/m2) Q3W during radiotherapy, then followed by adjuvant therapy with tislelizumab (200 mg) Q3W for 13 cycles. The primary endpoint was 2-year progression-free survival (PFS). The secondary endpoint included objective response rate (ORR), overall survival (OS) and safety. Results A total of 55 patients were enrolled from Sep. 2021 to Jun. 2023 with a median age of 43 (22-66) yrs and 76.4% (n = 42) of male. T4 account for 43.6% and N3 account for 67.3%. After a median follow-up of 16 months, 7 patients experienced disease progression with one patient in local relapse, three patients in regional relapse and three patients in distant metastasis. One-year PFS rate was 89.7%, and 1-year OS rate was 100%. According to RECIST1.1, 48 (87.3%) patients had objective response to neoadjuvant treatment, including 9 (16.4%) patient with CR, 39 (70.9%) patients with PR. The other 7 (12.7%) patients had SDa with a definition of tumor shrinkage occurred and below a 30% decrease in the sum of diameters of target lesions, taking as reference the baseline sum diameters. Throughout the neoadjuvant phase, 23 (41.8%) patients experienced grade 3-4 treatment-related adverse events (TRAE), with the most common being nausea (9[16.4%]) and leukopenia (8[14.5%]). Grade 3-4 TRAE appeared in 15 (27.3%) patients during concurrent chemoradiotherapy, with the most common being leukopenia (7[12.7%]) and mucositis (5[9.1%]). Conclusion For stage IVA nasopharyngeal carcinoma patients, neoadjuvant with tislelizumab plus GP and adjuvant tislelizumab treatment is safe and promising. Further follow-up is needed to confirm the long-term efficacy.

Abstract Background Easy-to-use, rapid, scalable, high-throughput, and cost-effective HPV tests are urgently needed for low-resource settings. Atila Biosystems’ high-throughput, cost-effective, and clinically validated ScreenFire HPV Risk Stratification (RS) assay identifies 13 high risk HPV (hrHPV) in 4 groups based on their oncogenic risk (i.e., HPV16, HPV18/45, HPV31/33/35/52/58, and HPV51/59/39/56/68). The current standard format is subject to laboratory contamination, which is common for any molecular PCR test. To overcome this drawback, Atila has recently upgraded it into an innovative, contamination-free Zebra BioDome format. The contamination-free feature makes this novel assay format more suitable for large-scale community- and population-based cervical screening. This study evaluated the analytical performance of the Zebra BioDome format. Methods We conducted a study to test the analytical performance of Zebra Biodome format in comparison to the results of using the ScreenFire HPV RS assay standard format on Biorad CFX-96 real-time PCR instrument. We used overall agreement rate and unweighted kappa value to compare the performance. Results The overall agreement for detection of hrHPV was 96.0% with unweighted kappa value 0.94 (95% confidence interval: 0.90–0.98). The agreement rates between hrHPV genotype 16 and risk stratification genotype group (HPV18/45, HPV31/33/35/52/58, and HPV51/59/39/56/68) were all > 97.5%. Conclusion The innovative ScreenFire HPV RS assay Zebra BioDome format produced highly concordant results with the standard format. The shared features by the two assay formats, such as easy-to-use, high throughput, cost-appropriate, and no requirements for DNA extraction. The unique contamination-prevention feature along with no requirement of preparation of reagents make the Zebra BioDome format more suitable for large-scale HPV screening to reduce global cervical cancer burden.

Background: MYD88 has been identified as a pivotal driver gene in hematologic B-cell malignancies. While MYD88 mutations are rare in western CLL populations (0.5%-3%), they are significantly more common in Asian cohorts, including China, Korea, India and Singapore, with an incidence of 10-20%. These mutations potentially correspond to a distinct CLL subtype with distinctive biological behaviors and survival outcomes. This study aims to delineate the clinical profiles and prognostic implications of MYD88-mutated (MYD88MUT) CLL patients under various treatment backgrounds. Methods: We analyzed clinical data from 1,081 CLL patients diagnosed between 2000 and 2024, who underwent MYD88 detection via next-generation sequencing or Sanger sequencing. We gathered information on laboratory indicators, IGHV status, genetic traits, treatment regimens, and survival rates. Results: In our cohort, we identified MYD88 mutations in 139 patients (12.86%). Specifically, 58 (39.7%) harboring the L265P variant, 49 cases (35.3%) with V217F, 15 (10.8%) with S219C, and 11 (7.2%) with M232T. MYD88MUT CLL showed a notable male predominance (P < 0.001) and a higher prevalence of IGHV mutations (95.0% vs. 60.6%, P < 0.001) compared to MYD88 wild-type (MYD88WT) patients. These patients also exhibited a higher presence of monoclonal gammopathy and marked absence of adverse cytogenetic aberrations, such as Del(17p) (1.6% vs. 11.9%, P < 0.001) and Del(11q) (1.6% vs. 12%, P < 0.001), with a reduced incidence of complex karyotypes (11.0% vs. 26.6%, P < 0.001). Similar proportions of atypical CLL with a Matutes score of 3 were observed between MYD88MUT and MYD88WT patients (10.1% vs. 8.0%, P=0.398). MYD88MUT CLL showed a reduced FMC7 negativity rate and a significantly diminished CD38 positivity. IGH VH3-7 and VH3-74 were prevalent in V217F and S219C mutations, whereas VH3-23 and VH4-34 were dominant in the MYD88 L265P variant. MYD88MUT CLL demonstrated superior clinical outcomes, including extended time to first treatment (TTFT), progression-free survival (PFS), and overall survival (OS) (m-TTFT 43 vs. 19 months, P < 0.001; m-PFS 132 vs. 49 months, P < 0.001; m-OS not reached vs. 135 months, P = 0.001). Even within the IGHV-mutated subpopulation, MYD88MUT CLL showed significantly improved outcomes compared to MYD88WT CLL (m-TTFT 47 vs. 32 months, P = 0.040; m-PFS 132 vs. 72 months, P < 0.001, m-OS not reached vs. 155 months, P = 0.019). MYD88MUT patients with IGHV-unmutated status demonstrated poorer outcomes, mirroring the outcomes of MYD88WT patients with unmutated IGHV. Patients were categorized into three treatment groups based on their first-line therapy: target therapy, immunotherapy, and chemotherapy. Subgroup analysis based on initial treatment showed that MYD88MUT patients experienced significantly enhanced PFS across all treatment modalities compared to non-mutated counterparts. In the MYD88WT cohort, patients undergoing target therapy had superior PFS and OS compared to immunotherapy and chemotherapy groups. However, in the MYD88MUT CLL group, all three treatment groups resulted in comparable PFS and OS, with impressive long-term outcomes across all groups (5-year PFS exceeding 90%, 5-year OS surpassing 80%, and 10-year OS above 70% in all three groups). The survival benefit of targeted therapy was not evident in MYD88MUT CLL. Among MYD88MUT patients, those harboring the L265P mutation (39.7%) experienced a notably shorter TTFT compared to non-L265P mutated patients. Nevertheless, there were no significant differences in PFS and OS based on the mutation sites. Conclusion: CLL patients with MYD88 mutations exhibited distinct biological characteristics, and had excellent survival outcomes, irrespective of the treatment modality.

Supplementary Figure 6. Survival outcomes for the entire cohort. (A) The progression free survival of the entire cohort; (B) The overall survival of the entire cohort.

Supplementary Table 6. The test performance of three methods in detection of CXCR4 mutations

Supplementary Table 2. Clinical characteristics and first-line treatment options for the entire symptomatic Waldenstrom macroglobulinemia (WM) cohort (n = 385) and for patients available for analysis of treatment response and outcome (n = 336).

Supplementary Table 3. The test performance of four methods in detection of MYD88 mutations.

Supplementary Table 5. Detection of MYD88 and CXCR4 mutation by different methods, NGS, next generation sequencing; AS-PCR, real-time

People living with the human immunodeficiency virus (HIV) are at a greater risk of developing hepatocellular carcinoma (HCC), potentially due to the stimulation of inflammation by HIV infection. Inflammation-related DNA methylation signatures obtained in liquid biopsy, such as circulating cell-free DNA (cfDNA), may serve as promising minimally invasive biomarkers that can inform diagnosis of HCC. Using data from 249 individuals with HIV (114 individuals with normal liver conditions, 69 with fibrosis, 30 with cirrhosis, and 36 with HCC), we constructed a cfDNA methylation-based inflammation score (inflammation-DNAm score) based on 54 CpGs previously associated with circulating C-reactive protein concentrations. Associations of DNAm scores with HCC were assessed using multivariable logistic regression models. Receiver operating characteristic analysis was conducted to assess the performance of discriminating HCC between the inflammation-DNAm score and alpha-fetoprotein (AFP), one of the current screening biomarkers. A higher inflammation-DNAm score was associated with a 29

Inetetamab is a neotype HER2-targeted monoclonal antibody with amino acids modified Fc segment which optimizes the antibody-dependent cellular cytotoxicity effect. However, robust evidence evaluating the combination of inetetamab combined with pertuzumab, paclitaxel and carboplatin (TCbIP) for neoadjuvant therapy is still lacking. This study aimed to evaluate the efficacy and safety of TCbIP as a neoadjuvant therapy for patients with locally advanced (LA) HER2-positive breast cancer. This phase II trial included female patients with histologically confirmed stage IIA to IIIC HER2-positive primary invasive breast cancer. Eligible patients received TCbIP treatment every three weeks for a maximum of six cycles followed by surgery. The primary endpoint was pathologic complete response (pCR, ypT0/is ypN0) rate. Key secondary endpoints included near pCR (npCR, residual breast disease <1cm) rate, objective response rate (ORR) and safety. From November 2021 to May 2023, 28 patients were enrolled in the trial. One patient received one cycle of the study treatment and was lost to follow-up without surgery, and four patients received one cycle of the study treatment and were still in treatment, leaving 23 patients in the ITT population. Among these 23 patients (82.6% in stage III), 16 patients completed the study treatment and surgery (PP population) and six patients were still undergoing neoadjuvant treatment. The ORR was 91.3% (21/23) in the ITT population and 93.8% (15/16) in the PP population. Among the 16 patients in the PP population, 10 patients (62.5%) achieved pCR. A total of 14 patients (87.5%) achieved npCR. For patients with hormone receptor (HR) negative and positive tumors, the pCR rates were 88.9% (8/9) and 28.6% (2/7), respectively. The most common grade 3 adverse event was neutropenia (25.0%). No significant reduction in the left ventricular ejection fraction was observed in any patient. Neoadjuvant therapy with TCbIP has shown promising efficacy and manageable toxicity in patients with HER2-positive LA breast cancer.

The phase Ⅲ CASPIAN study established durvalumab (D) plus etoposide with cisplatin or carboplatin (EP) as first-line standard of care (1L SoC) of ES-SCLC. The multicenter, single-arm, phase Ⅲb ORIENTAL study evaluated 1L D+EP in a large, real-world-like patient (pt) cohort in China. Preliminary safety and efficacy results were consistent with CASPIAN. Here we report final results and subgroup analysis.