AbstractDifferences in the cervicovaginal microbiome may influence the persistence of HPV and therefore, the progression to cervical cancer. We aimed to analyze and compare the metatranscriptome of cervical cancers positive for HPV 16 and 18 with those positive for other HPV types to understand the microbiome’s influence on oncogenicity.

摘要宫颈阴道微生物组的差异可能会影响HPV的持久性，从而影响宫颈癌的进展。我们旨在分析和比较HPV 16和18阳性的宫颈癌与其他HPV类型阳性的宫颈癌的转录组，以了解微生物组对致癌性的影响。

RNA sequencing data from a total of 222 invasive cervical cancer cases (HPV16/18 positive (n=42) and HPV “Other types” (n=180)) were subjected to taxonomy classification (Kraken 2) including bacteria, virus and fungi to the level of species. With a median depth of 288,080.5 reads per sample, up to 107 species (38 bacterial, 16 viral and 53 fungal) were identified.

对来自222例浸润性宫颈癌病例（HPV16/18阳性（n=42）和HPV“其他类型”（n=180））的RNA测序数据进行分类学分类（Kraken 2），包括细菌，病毒和真菌到物种水平。每个样品的中位深度为288080.5读数，鉴定出多达107种（38种细菌，16种病毒和53种真菌）。

Diversity analyses revealed no significant differences in viral or fungal species between HPV16/18 and other HPV types. Bacterial alpha diversity was significantly higher in the 'Other HPV types' group for the Observed index (p=0.0074) (but not for Shannon). Cumulative species curves revealed greater species diversity in the “Other HPV types” group compared to “HPV16/18 but no significant differences in species abundance were found between HPV groups.

多样性分析显示，HPV16/18与其他HPV类型之间的病毒或真菌种类没有显着差异。在观察到的指数中，“其他HPV类型”组的细菌α多样性显着更高（p=0.0074）（但香农没有）。累积物种曲线显示，“其他HPV类型”组的物种多样性高于“HPV16/18”，但HPV组之间的物种丰度没有显着差异。

The study did not detect strong significant microbiome differences between HPV 16/18 and other HPV types in cervical cancers. Further research is necessary to explore potential factors influencing the oncogenicity of different HPV types and their interaction with the cervical microbiome..

该研究未发现HPV 16/18与宫颈癌中其他HPV类型之间存在显着的微生物组差异。有必要进一步研究以探索影响不同HPV类型致癌性的潜在因素及其与宫颈微生物组的相互作用。。

IntroductionHuman papillomaviruses (HPVs) are a diverse group of double-stranded DNA viruses comprising up to 225 different types, with new types continuously being identified1,2,3,4. Among these, approximately 12 HPV types are classified as oncogenic, high-risk HPV genotypes, with persistent infection by these types being a necessary cause for cervical cancer.The oncogenic potential of different high-risk HPV types varies, existing profound differences in carcinogenicity among the HPV types.

引言人乳头瘤病毒（HPV）是一组多样的双链DNA病毒，包括多达225种不同的类型，不断发现新的类型1,2,3,4。其中，大约12种HPV类型被归类为致癌的高危HPV基因型，这些类型的持续感染是宫颈癌的必要原因。不同高危型HPV的致癌潜力各不相同，HPV类型之间的致癌性存在深刻差异。

HPV 16 has by far the highest oncogenic potential, causing more than half of cervical cancers (62.4%), followed by HPV 18 (15.3%)5. Besides HPV 16 and 18, there are an additional 5 types (HPV 31, 33, 35, 45, 52 and 58) that are found in >2% of cervical cancers and jointly account for an additional 20% of cervical cancers5,6,7,8.

到目前为止，HPV 16具有最高的致癌潜力，导致超过一半的宫颈癌（62.4%），其次是HPV 18（15.3%）5。除HPV 16和18外，还有5种类型（HPV 31,33,35,45,52和58）在>2%的宫颈癌中发现，共同占宫颈癌的20%5,6,7,8。

The least carcinogenic types (HPV 39, 51, 56 and 59) each contribute less than 1% of cervical cancer cases5.These strong differences in carcinogenicity are the reason why some HPV tests analyze for HPV 16 and HPV 18 separately (“high-risk” types) and report an aggregated result “Other HPV” for some other HPV types (oncogenic, probably oncogenic and possibly oncogenic)5.

致癌性最低的类型（HPV 39,51,56和59）各自占宫颈癌病例的不到1%。这些致癌性的强烈差异是一些HPV检测分别分析HPV 16和HPV 18的原因（“高风险”类型）并报告其他一些HPV类型（致癌，可能致癌和可能致癌）的“其他HPV”汇总结果5。

Mechanisms on why some HPV types are more oncogenic than others remain not fully understood. It is well known that persistence of the virus is crucial for carcinogenesis, and several authors have reported that persistence can be favored by chronic inflammation in the tissue caused by an imbalanced cervicovaginal microbiome9,10.

为什么一些HPV类型比其他类型更具致癌性的机制尚不完全清楚。众所周知，病毒的持久性对于致癌作用至关重要，一些作者报道，由不平衡的宫颈阴道微生物组引起的组织慢性炎症可能有利于持久性[9,10]。

A loss of Lactobacillus genera can lead to the colonization of anaerobic opportunistic bacteria inducing pro-inflammatory cytokine and ROS production, as an example11,12. All this evidence positioned microbiome profiles as good candidates to understand the underlying .

例如，乳酸杆菌属的丧失可导致厌氧机会性细菌的定植，诱导促炎细胞因子和ROS产生[11,12]。所有这些证据都将微生物组概况定位为了解基础的良好候选者。

Data availability

数据可用性

All sequencing files (non-human sequences) used in the present study are publicly available at the Sequence Read Archive (SRA) within the bio-project ID PRJNA563802.

本研究中使用的所有测序文件（非人类序列）均可在生物项目ID PRJNA563802内的序列读取档案（SRA）中公开获得。

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Download referencesAcknowledgementsAuthors would also like to thank Head of Department Joakim Dillner for continuous encouragement and support.FundingThis Project was funded by the Human Exposome Assessment Platform (Project No. 874662) granted by Horizon 2020.Open access funding provided by Karolinska Institute.Author informationAuthor notesThese authors contributed equally: Jiayao Lei and Laila Sara Arroyo Mühr.Authors and AffiliationsDepartment of Genomic Medicine, African Institute of Biomedical Science and Technology, 911 Boronia Township, Beatrice, Harare, ZimbabweMaire Hidjo & Collen MasimirembwaUniversity of Witwatersrand Sydney Brenner Institute for Molecular Biosciences, Johannesburg, 2193, South AfricaMaire Hidjo & Collen MasimirembwaCenter for Cervical Cancer Elimination, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, 141 86, Stockholm, SwedenDhananjay Mukhedkar, Jiayao Lei & Laila Sara Arroyo MührHopsworks AB, Åsögatan 119, Plan 2, 116 24, Stockholm, SwedenDhananjay MukhedkarDepartment of Medical Epidemiology and Biostatistics, Karolinska Institutet, 171 77, Solna, SwedenJiayao LeiAuthorsMaire HidjoView author publicationsYou can also search for this author in.

下载参考文献致谢作者还要感谢部门负责人JoakimDillner的不断鼓励和支持。资助该项目由Horizon 2020授予的人类暴露评估平台（项目编号874662）资助。开放获取资金由卡罗林斯卡研究所提供。作者信息作者注意到这些作者做出了同样的贡献：Jiayao Lei和Laila Sara Arroyo Mühr。作者和附属机构非洲生物医学科学与技术研究所基因组医学系，911 Boronia Township，Beatrice，Harare，ZimbabweMaire Hidjo＆Collen MasimirembwaUniversity of Witwatersrand Sydney Brenner Institute for Molecular Biosciences，约翰内斯堡，2193，南非Maire Hidjo＆Collen MasimirembwaCenter for Cervical Cancer Elimination，Department of Clinical Science，Intervention and Technology（CLINTEC），Karolinska Institutet，141 86，Stockholm，Swedhanjay Mukhedkar，Jiayao Lei＆Laila SaraArroyo MührHopsworks AB An 119，Plan211624，Stockholm，SwedenDhananjay MukhedkardDepartment of Medical Epidemiology and Biostatistics，Karolinska Institutet，171 77，Solna，SwedenJiayao LeiAuthorsMaire HidjoView author Publications您也可以在中搜索这位作者。

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PubMed Google ScholarContributionsMarie Hidjo: Data curation, Formal analysis, Methodology, Validation, Writing – original draft preparation. Dhananjay Mukhedkar: Formal analysis, Methodology, Validation, Writing – review and editing. Collen Masimirembwa: Supervision, Validation, Writing – review and editing.

PubMed Google ScholarContributionsMarie Hidjo：数据管理，正式分析，方法论，验证，写作-原始草案准备。Dhananjay Mukhedkar：形式分析，方法论，验证，写作-评论和编辑。科伦·马西米尔·姆布瓦（CollenMasimirembwa）：监督、验证、写作——审查和编辑。

Jiayao Lei: Conceptualization, Investigation, Methodology, Project administration, Supervision, Validation, Writing – review and editing. Laila Sara Arroyo Mühr: Conceptualization, Investigation, Methodology, Project administration, Supervision, Validation, Writing – original draft preparation, Writing – review and editing.

雷家耀：概念化，调查，方法论，项目管理，监督，验证，写作-审查和编辑。Laila Sara Arroyo Mühr：概念化，调查，方法论，项目管理，监督，验证，写作-原稿准备，写作-审查和编辑。

All the authors have read and approved the final manuscript.Corresponding authorCorrespondence to.

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The authors declare no competing interests.

作者声明没有利益冲突。

Ethics approval and consent to participate

道德批准和同意参与

Ethical approval was granted to collect archival material from all cervical cancer cases, to perform histopathology review of the diagnostic slides, and to collect the formalin-fixed paraffin-embedded (FFPE)-blocks for HPV-genotyping. The Swedish Ethical Review Board Authority of Stockholm determined that, due to the population-based nature of the study, informed consent from study participants was not required (EPN-Dnr: 2011/1026-31/4) and collection of the samples for histology review and HPV-typing was also allowed (EPN-Dnr: 2012/1028/32)..

获得了伦理批准，可以从所有宫颈癌病例中收集档案材料，对诊断载玻片进行组织病理学检查，并收集福尔马林固定石蜡包埋（FFPE）块进行HPV基因分型。斯德哥尔摩瑞典伦理审查委员会管理局确定，由于该研究以人群为基础，不需要研究参与者的知情同意（EPN Dnr:2011/1026-31/4），也允许收集样本进行组织学检查和HPV分型（EPN Dnr:2012/1028/32）。。

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Reprints and permissionsAbout this articleCite this articleHidjo, M., Mukhedkar, D., Masimirembwa, C. et al. Cervical cancer microbiome analysis: comparing HPV 16 and 18 with other HPV types.

转载和许可本文引用本文Hidjo，M.，Mukhedkar，D.，Masimirembwa，C。等人。宫颈癌微生物组分析：比较HPV 16和18与其他HPV类型。

Sci Rep 14, 22014 (2024). https://doi.org/10.1038/s41598-024-73317-8Download citationReceived: 11 June 2024Accepted: 16 September 2024Published: 24 September 2024DOI: https://doi.org/10.1038/s41598-024-73317-8Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

科学报告1422014（2024）。https://doi.org/10.1038/s41598-024-73317-8Download引文接收日期：2024年6月11日接受日期：2024年9月16日发布日期：2024年9月24日OI：https://doi.org/10.1038/s41598-024-73317-8Share本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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KeywordsCervical cancerHuman papillomavirusCervical microbiomeMetatranscriptome

关键词宫颈癌人类乳头瘤病毒宫颈微生物组转录组

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