Bingjian Lu

An immunosuppressive tertiary lymphoid structure is associated with adverse prognosis in gastric-type endocervical adenocarcinoma

Abstract Background Gastric-type adenocarcinoma (GAC) is the predominant subtype of HPV-independent endocervical adenocarcinoma, characterized by aggressive clinical behavior and poor prognosis. However, its tumor immune microenvironment (TIME) remains largely unexplored. Methods We systematically profiled the immune landscape using bulk RNA sequencing and multiplex immunofluorescence staining and comprehensively analyzed the associations between immune features, clinicopathological parameters, and patient outcomes in a multi-institutional cohort of 153 GAC cases. Results GAC tumors exhibited sparse T cell but abundant macrophage infiltration, predominantly with M2-polarized macrophages in advanced stage. Tumors with tertiary lymphoid structure (TLS) demonstrated an immunosuppressive milieu characterized by enrichment of B cells, PD1-CD8+ T cells, and regulatory T cells. While TLS-positive patients were associated with poorer prognosis, 2 patients showed improved survival with chemoradiotherapy. Moreover, the TLS and FIGO stage-based prognostic model was robust in the risk stratification for tumor recurrence. Conclusions We reveal a distinct immunosuppressive TIME and TLS in GAC. TLS might confer an adverse prognosis. Our findings provide valuable clues for individualized therapy for this aggressive cancer in future.

Endocervical adenocarcinoma with a micropapillary component: a clinicopathologic analysis in the setting of current WHO classification

Our study aimed to investigate the clinicopathologic and molecular features of endocervical adenocarcinoma with a micropapillary component (EAC-MP) in the setting of current classification schema. We investigated 26 EAC-MP from consecutive 511 adenocarcinomas. HER2 status was analyzed by immunohistochemistry and fluorescence in situ hybridization. Four cases were performed with targeted next-generation sequencing (NGS). We found that HPV-associated adenocarcinomas (HPVA) with a micropapillary component (HPVA-MP) (n = 12) had a higher frequency of large tumor size (> 2 cm), Silva pattern C (12/12, 100%), invasion of the deep cervical wall (> 2/3) (8/12, 66.7%), lymphovascular space invasion (LVSI) (11/12, 91.7%), lymph node metastasis (4/11, 36.4%), FIGO stage III/IV (4/12, 33.3%), and HER2 amplification (3/12, 25%, P = 0.015), compared to those without (HPVA-NMP (all P  0.05). NGS identified significant mutations in STK11, TERT, ERBB2, TP53, PIK3CA, ARID1A, and NTRK2. We conclude that the micropapillary structure is an indicator for unfavorable clinical outcomes in HPVA, and can aid in the prognostic stratification of Silva pattern C EAC. The presence of HER2 amplification and specific gene mutations raise the possibility for targeted therapy in the future.

Uterine fibrosarcomatous dermatofibrosarcoma protuberans (FS-DFSP) with COL1A1::PDGFB fusion and concurrent TP53/ERBB2 mutations: Case report and molecular characterization

Uterine dermatofibrosarcoma protuberans (DFSP) with COL1A1::PDGFB fusion is an exceptionally rare sarcoma, and its high-grade variant remains poorly characterized. We reported a 39-year-old Chinese woman with a high-grade uterine sarcoma exhibiting morphological features of fibrosarcomatous DFSP (FS-DFSP), including spindle cells arranged in storiform and herringbone patterns, focal myoid/epithelioid differentiation, and myxoid changes. Immunohistochemistry revealed variable CD34 expression, focal desmin/SMA positivity, and aberrant p53 overexpression. Targeted next-generation sequencing identified a COL1A1-PDGFB (C5:P2) fusion, a pathogenic TP53 (c.832 C>G; p.P278A) and ERBB2 (c.2524 G>A; p.V842I) mutation. Dual-color fluorescence in situ hybridization confirmed COL1A1-PDGFB fusion. This represented the first report of high-grade uterine DFSP with concurrent TP53 and ERBB2 mutations, expanding the molecular spectrum of this entity and suggesting potential therapeutic implications for dual PDGFR/HER2 inhibition in aggressive cases.

Further confirmation of a highly prognostic grading scheme based upon tumour budding and cell cluster size in cervical squamous cell carcinoma

AimsOur study aimed to further confirm the clinical significance of the tumour budding activity and cell nest size‐based (TBNS) grading scheme in cervical squamous cell carcinomas (SCC).Methods and resultsWe applied the TBNS system to assess the prognostic value in an institutional cohort of well‐annotated cervical SCC consisting of 312 consecutive cases with surgical resection, no neoadjuvant chemotherapy and higher than stage pT1a. We found that high budding activity, single cell and TBNS grade 3 were more frequently associated with a decreased overall survival (OS) time and disease‐free survival (DFS) time (P < 0.001) and several other clinicopathological factors, including lymphovascular space invasion, lymph node metastasis, advanced Federation of Gynecology and Obstetrics (FIGO) stage and deep invasion of the cervical wall (> 2/3) (P < 0.05). On multivariate analysis, TBNS grade 3 was an adverse indicator for OS and DFS independently of age, invasion of the cervical wall and FIGO stage (P < 0.05). By comparison, the conventional three‐tiered grading system was not associated with OS and DFS in cervical SCC (P > 0.05).ConclusionsOur study further confirms that the TBNS grading scheme is robust in prognostic assessment in cervical SCC that outperforms the conventional three‐tiered grading system. It is applicable to add TBNS grade into routine diagnostic practice.

Human epidermal growth factor 2 (HER2) amplification in uterine serous carcinoma: an analysis of prognosis and immune microenvironment

Uterine serous carcinoma (USC) is a biologically aggressive subtype of endometrial cancer. Anti-human epidermal growth factor receptor 2 (HER2) therapy has demonstrated its promising effects on HER2-positive USC. However, data on prognostic relevance and immune microenvironment are limited in HER2-positive USC. This study aimed to determine the clinicopathologic features, prognosis, and the immune microenvironment trait in HER2 status in USC. We applied immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and multi-color immunofluorescence to investigate HER2 expression and amplification, PD-L1 expression, and tumor infiltration lymphocytes (TIL) in 77 USC (61 pure and 16 mixed-type USC). HER2 IHC 1 + , 2 + , and 3 + were found in 26, 18, and 10 USC, respectively. HER2 staining frequently had an incomplete membrane (basolateral or "U"-shaped) pattern. Twenty-three cases (23/54, 42.6%) showed an intra-tumor heterogeneous staining. HER2 amplification was present in 16/77 (20.8%) USC. HER2 amplification was significantly associated with deep myometrial invasion (> 1/2), and increased intra-epithelial and stromal density of CD20 + or CD8 + TIL (all P  0.05). HER2 amplification was associated with poor overall and progression-free survival in USC, but lost the prognostic significance on multivariate analysis. We concluded that HER2 amplified USC had adverse clinical outcomes, but showed the potential active immune microenvironment. Our findings raised the possibility of the combined anti-HER2 and immunotherapy for HER2-positive USC in the future.

Retrospective analysis of cytology and high‐risk HPV testing in 1067 endocervical adenocarcinomas and precursor lesions

AbstractBackgroundCytology and high‐risk human papilloma virus (hrHPV) cotesting is the mainstay in the detection of cervical carcinoma.MethodsEndocervical adenocarcinoma (EAC) is divided into HPV‐associated adenocarcinoma (HPVA) and HPV‐independent adenocarcinoma (HPVI) by the World Health Organization classification (2020). The detection effect of cotesting is suggested to be different among EAC subtypes and precursors, but has not well‐documented yet. In this study, the authors retrospectively analyzed cotesting among adenocarcinoma in situ (AIS), HPVA, and HPVI. The cohort included 569 AIS and 498 EAC consisting of 371 (74.5%) HPVA, 111 (22.3%) HPVI, and 16 (3.2%) adenocarcinoma, not otherwise specified.ResultsThe authors found that AIS patients were significantly younger than HPVA and HPVI (mean ± SD, years: 40.7 ± 8.6; HPVA, 44.8 ± 9.3; HPVI, 50.0 ± 11.3; p < .001) and had a higher prevalence of concurrent squamous intraepithelial lesions (75.5%, HPVA, 37.2%; HPVI, 12.6%; p < .001). The detection rate of hrHPV test or cytology was substantially higher in AIS and HPVA than in HPVI (97.7% and 90.2% vs. 16.5%, p < .001, or 71.1% and 71.9% vs. 60.7%, p = .042, respectively). Cytology and hrHPV cotesting was superior to a single test in the detection of EAC and AIS. The detection rate of cotesting amounted to 100% in AIS and 94.3% in HPVA but was substantially lower in HPVI (72.2%) (p < .001).ConclusionsThe authors conclude that cytology and hrHPV cotesting can maximize the detection effect for HPVA and AIS but is not optimal for HPVI.

A clinicopathological and molecular analysis of cervical carcinomas with basaloid features

AimsTo investigate the relationship between adenoid basal carcinoma (ABC), adenoid cystic carcinoma (ACC) and squamous cell carcinoma (SCC) in the uterine cervix.Methods and resultsWe analysed the clinicopathological and molecular features in two pure ABCs, 15 SCCs with ABC‐/ACC‐like features and seven basaloid SCCs (BSCCs) by chart review, immunohistochemistry, human papillomavirus (HPV) RNA in‐situ hybridisation and fluorescence in‐situ hybridisation. All patients were alive with no evidence of disease, except for one patient with ACC‐like features who died of disease at 18 months post diagnosis. The mixed carcinomas comprised variable SCCs and ABC‐/ACC‐like components displaying vague transitional zones. All components consistently showed diffuse p16, p63 and SOX2, variable cytokeratin (CK)7 and CK17 and rare Ber‐EP4 and MYB expression; there was a substantially lower Ki67 index in pure ABCs and the ABC‐like components. The ACC‐like components showed no myoepithelial differentiation (SMA, calponin and S100) and MYB gene fusions. CK7, CK17 and Ber‐EP4 were characteristically stronger in BSCCs than in the mixed carcinomas (P < 0.01). High‐risk HPV (HR‐HPV) E6/E7 mRNA was detected in 12 mixed carcinomas and seven BSCCs, but not in pure ABCs. The HR‐HPV mRNA expression was higher in the SCC components and BSCCs than in the ABC‐like components of mixed carcinomas (P < 0.05).ConclusionsThe ACC‐like components in mixed carcinomas probably represent the morphological mimics of salivary ACCs. ABC‐like components may be the potential precursor of the ACC‐like and SCC components. HR‐HPV oncogenes may play a role in the pathogenesis of SCCs with ABC‐/ACC‐like features.

A novel tumor budding and cell nest size-based grading system outperforms conventional methods in vulvar squamous cell carcinoma

The tumor budding and tumor cell nest size-based (TBNS) grading scheme is an emerging prognostic indicator for squamous cell carcinoma (SCC) across various organs; however, its significance in vulvar SCC (VSCC) remains poorly investigated. In this study, we applied the TBNS grading system to an institutional cohort of 62 consecutive surgically resected VSCC cases (39 HPV-independent and 23 HPV-associated), excluding patients with neoadjuvant chemotherapy or FIGO stage IA disease. High tumor budding activity, small cell nest size, and high TBNS grade were significantly associated with reduced overall survival (OS) and disease-free survival (DFS) in VSCC, as well as with adverse clinicopathologic features such as lymphovascular space invasion, perineural involvement, lymph node metastasis, and advanced FIGO stage (p  0.05). We conclude that the TBNS grading system is a promising prognostic indicator for VSCC, outperforming conventional grading systems. Further validation in larger cohorts is needed to expand the clinical applicability of this grading system in VSCC.

Grading of endocervical adenocarcinoma: a novel prognostic system based on tumor budding and cell cluster size

A novel 3-tiered grading system based on tumor budding activity and cell nest size has been validated to be highly prognostic in organ-wide squamous cell carcinomas. In this study, we applied a similar grading system with slight modification to assess the prognostic value in an institutional cohort of well annotated endocervical adenocarcinomas (EAC) consisting of 398 consecutive cases with surgical resection, no neoadjuvant chemotherapy, and higher than stage pT1a. Each case was reviewed by the International Endocervical Adenocarcinoma Criteria and Classification (IECC) and Silva pattern classification, and scored on tumor budding activity and cell cluster size to form the basis of a novel grading system. High budding activity, small tumor cell cluster size, and novel grade 3 were more frequently associated with a decreased overall survival time and tumor recurrence time (p  0.05). In conclusion, our study demonstrates that the grading system based on tumor budding activity and cell cluster size is robust in prognostic assessment that outperforms the conventional FIGO grading and Silva pattern classification in EAC. The novel grading system, if further validated, could be applicable in routine pathologic descriptions of EAC by providing useful information in clinical decision-making.

LncRNA SFTA1P promotes cervical cancer progression by interaction with PTBP1 to facilitate TPM4 mRNA degradation

AbstractLong non-coding RNAs (lncRNAs) play key roles in cancer development and progression. However, the biological function and clinical significance of most lncRNAs in cervical cancer remain elusive. In this study, we explore the function and mechanism of lncRNA surfactant associated 1 (SFTA1P) in cervical cancer. We firstly identified SFTA1P by analyzing the RNA sequencing data of cervical cancer from our previous study and from The Cancer Genome Atlas (TCGA). We then verified SFTA1P expression by qRT-PCR. The cell proliferation and migration capacity of SFTA1P was assessed by using CCK-8, colony formation, transwell and wound healing assays. RNA pull-down, RNA immunoprecipitation (RIP), RNA stability and western blot assays were used to reveal potential mechanisms. Athymic nude mice were used to evaluate tumorigenicity and metastasis in vivo. SFTA1P is upregulated in cervical tumor tissues and its high expression is associated with poor prognosis. Biologically, knockdown of SFTA1P inhibited the proliferation, migration, and invasion of cervical cancer cells in vitro, as well as tumorigenesis and metastasis in vivo. Mechanistically, SFTA1P was shown to interact with polypyrimidine tract binding protein 1 (PTBP1) to regulate the stability of tropomyosin 4 (TPM4) mRNA, thereby resulting in malignant cell phenotypes. TPM4 knockdown could attenuate the suppression of cell progression induced by either SFTA1P or PTBP1 knockdown. Our findings demonstrate that SFTA1P can promote tumor progression by mediating the degradation of TPM4 mRNA through its interaction with PTBP1 protein. This provides a potential therapeutic strategy to target the SFTA1P-PTBP1-TPM4 axis in cervical cancer.

A Retrospective Analysis of 1151 Chinese Women With Atypical Glandular Cells: Impact of High‐Risk Human Papilloma Virus Status and Age on Cervical and Endometrial Neoplasia Risk

ABSTRACTObjectiveTo investigate the clinical significance of high‐risk human papillomavirus (hrHPV) testing and age in women with atypical glandular cells (AGC) on Pap tests.MethodsWe retrospectively analyzed the relationship between hrHPV and cytology co‐testing, follow‐up histopathology within 6 months, age, and other demographic features in a large cohort of AGC patients from a single Chinese academic institution.ResultsAmong1510 AGC patients, 1151, 1260, and 866 patients had histopathologic results, hrHPV co‐testing, and both, respectively. High‐grade squamous intraepithelial lesions (HSIL)/adenocarcinoma in situ (AIS) and worse lesions (HSIL/AIS+) and endometrial atypical hyperplasia (EAH)/endometrial carcinoma (EC) (EAH/EC+) were present in 27.8% (281/1009) and 25.1% (113/450) patients, respectively. hrHPV was positive in 34.5% (435/1260) patients. Among hrHPV+ patients with genotyping by the Aptima assay, 48.8% (125/256) had HPV16 and/or 18/45. Compared to hrHPV− patients, hrHPV+ women had a significantly increased risk of HSIL/AIS+ (OR: 10.958, 95% CI: 7.657–15.682), particularly with HPV16 and/or18/45 (OR: 3.556, 95% CI: 2.122–5.959; both p < 0.001). Among hrHPV− patients, those aged ≥ 40 years had higher risks of HSIL/AIS+ (including 18 HPV‐independent and 11 metastatic adenocarcinomas; OR: 5.882, 95% CI: 2.702–12.806) and EAH/EC+ (OR: 7.622, 95% CI: 3.188–18.225) compared to younger patients (both p < 0.001).ConclusionsCombining hrHPV testing and age effectively stratifies risks in AGC patients: hrHPV positivity predicts high‐grade cervical lesions, while older hrHPV‐negative women are more likely to have endometrial or extra‐uterine malignancies. These findings may optimize management of AGC patients with cumulative supportive data.