Patient-derived and mouse endo-ectocervical organoid generation, genetic manipulation and applications to model infection

Rajendra Kumar Gurumurthy, Stefanie Koster, Naveen Kumar, Thomas F. Meyer, Cindrilla Chumduri

Published: 2022-05-10 DOI: 10.1038/s41596-022-00695-6

Organoids

Cervical Cancer

Genetic Manipulation

Infection Modeling

Stem Cells

AI 解读

Fabrication of angstrom-scale two-dimensional channels for mass transport

Efficient and strand-specific profiling of replicating chromatin with enrichment and sequencing of protein-associated nascent DNA in mammalian cells

The eINTACT method for studying nuclear changes in host plant cells targeted by bacterial effectors in native infection contexts

Rapid reaction optimization by robust and economical quantitative benchtop19F NMR spectroscopy

Profiling native pulmonary basement membrane stiffness using atomic force microscopy

Photoimmunotechnology as a powerful biological tool for molecular-based elimination of target cells and microbes, including bacteria, fungi and viruses

Measuring key human carbohydrate digestive enzyme activities using high-performance anion-exchange chromatography with pulsed amperometric detection

Neural cell isolation from adult macaques for high-throughput analyses and neurosphere cultures

Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds

Neuronal subtype-specific growth cone and soma purification from mammalian CNS via fractionation and fluorescent sorting for subcellular analyses and spatial mapping of local transcriptomes and proteomes

查看全部

Sections

Figures

References

The cervix is the gateway to the upper female reproductive tract, connecting the uterus and vagina. It plays crucial roles in fertility and pregnancy maintenance from onset until delivery of the fetus, and prevents pathogen ascension. Compromised function
Introduction
Materials
Reagents
Equipment
Reagent setup
Procedures
Troubleshooting
Timing
Anticipated results
- References
References
ACKNOWLEDGMENTS
ACKNOWLEDGMENTS

Chumduri, C. & Turco, M. Y. Organoids of the female reproductive tract. J. Mol. Med. 99, 531–553 (2021).
Wira, C. R., Rodriguez-Garcia, M. & Patel, M. V. The role of sex hormones in immune protection of the female reproductive tract. Nat. Rev. Immunol. 15, 217–230 (2015).
Wira, C. R., Grant-Tschudy, K. S. & Crane-Godreau, M. A. Epithelial cells in the female reproductive tract: a central role as sentinels of immune protection. Am. J. Reprod. Immunol. 53, 65–76 (2005).
Chumduri, C. et al. Opposing Wnt signals regulate cervical squamocolumnar homeostasis and emergence of metaplasia. Nat. Cell Biol. 23, 184–197 (2021).
Martyn, F., McAuliffe, F. M. & Wingfield, M. The role of the cervix in fertility: is it time for a reappraisal? Hum. Reprod. 29, 2092–2098 (2014).
WCRF (2018).
Chan, C. K., Aimagambetova, G., Ukybassova, T., Kongrtay, K. & Azizan, A. Human papillomavirus infection and cervical cancer: epidemiology, screening, and vaccination-review of current perspectives. J. Oncol. 2019, 3257939 (2019).
Zhu, H., Shen, Z., Luo, H., Zhang, W. & Zhu, X. Chlamydia trachomatis infection-associated risk of cervical cancer: a meta-analysis. Medicine 95, e3077 (2016).
Shen, R., Richter, H. E. & Smith, P. D. Interactions between HIV-1 and mucosal cells in the female reproductive tract. Am. J. Reprod. Immunol. 71, 608–617 (2014).
WHO (2018).
Schutgens, F. & Clevers, H. Human organoids: tools for understanding biology and treating diseases. Annu. Rev. Pathol. 15, 211–234 (2020).
Kim, J., Koo, B. K. & Knoblich, J. A. Human organoids: model systems for human biology and medicine. Nat. Rev. Mol. Cell Biol. 21, 571–584 (2020).
Koster, S. et al. Modelling Chlamydia and HPV co-infection in patient-derived ectocervix organoids reveals distinct cellular reprogramming. Nat. Commun. 13, 1030 (2022).
Clevers, H., Loh, K. M. & Nusse, R. Stem cell signaling. An integral program for tissue renewal and regeneration: Wnt signaling and stem cell control. Science 346, 1248012 (2014).
Hsu, Y. C., Li, L. & Fuchs, E. Transit-amplifying cells orchestrate stem cell activity and tissue regeneration. Cell 157, 935–949 (2014).
Huch, M. et al. Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell 160, 299–312 (2015).
Rheinwald, J. G. & Green, H. Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes. Nature 265, 421–424 (1977).
Llames, S., García-Pérez, E., Meana, Á., Larcher, F. & del Río, M. Feeder layer cell actions and applications. Tissue Eng. B 21, 345–353 (2015).
Herfs, M. et al. A discrete population of squamocolumnar junction cells implicated in the pathogenesis of cervical cancer. Proc. Natl Acad. Sci. USA 109, 10516–10521 (2012).
Koskela, P. et al. Chlamydia trachomatis infection as a risk factor for invasive cervical cancer. Int. J. Cancer 85, 35–39 (2000).
Ssedyabane, F. et al. HPV-chlamydial coinfection, prevalence, and association with cervical intraepithelial lesions: a pilot study at Mbarara Regional Referral Hospital. J. Cancer Epidemiol. 2019, 9092565 (2019).
Lohmussaar, K. et al. Patient-derived organoids model cervical tissue dynamics and viral oncogenesis in cervical cancer. Cell Stem Cell 28, 1380–1396 (2021).
Hoffmann, K. et al. Stable expansion of high-grade serous ovarian cancer organoids requires a low-Wnt environment. EMBO J. 39, e104013 (2020).
Lohmussaar, K. et al. Assessing the origin of high-grade serous ovarian cancer using CRISPR-modification of mouse organoids. Nat. Commun. 11, 2660 (2020).
Dolat, L. & Valdivia, R.H. An endometrial organoid model of interactions between Chlamydia and epithelial and immune cells. J. Cell Sci. 134, jcs252403 (2021).
Kessler, M. et al. Chronic Chlamydia infection in human organoids increases stemness and promotes age-dependent CpG methylation. Nat. Commun. 10, 1194 (2019).
Fatehullah, A., Tan, S. H. & Barker, N. Organoids as an in vitro model of human development and disease. Nat. Cell Biol. 18, 246–254 (2016).
Sato, T. et al. Single Lgr5 stem cells build crypt–villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009).
Katz, D. F. Human cervical mucus: research update. Am. J. Obstet. Gynecol. 165, 1984–1986 (1991).
Merbah, M. et al. Cervico-vaginal tissue ex vivo as a model to study early events in HIV-1 infection. Am. J. Reprod. Immunol. 65, 268–278 (2011).
Michelini, M., Rosellini, A., Simoncini, T., Papini, S. & Revoltella, R. P. A three-dimensional organotypic culture of the human uterine exocervix for studying mucosal epithelial differentiation and migrating leukocytes. Differentiation 72, 138–149 (2004).
Cattaneo, C. M. et al. Tumor organoid-T-cell coculture systems. Nat. Protoc. 15, 15–39 (2020).
Zadora, P. K. et al. Integrated phosphoproteome and transcriptome analysis reveals chlamydia-induced epithelial-to-mesenchymal transition in host cells. Cell Rep. 26, 1286–1302 (2019).
Farin, H. F., Van, Es,J. H. & Clevers, H. Redundant sources of Wnt regulate intestinal stem cells and promote formation of Paneth cells. Gastroenterology 143, 1518–152s (2012).
Willert, K. et al. Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature 423, 448–452 (2003).
Gurumurthy, R. K. et al. A loss-of-function screen reveals Ras-and Raf-independent MEK–ERK signaling during Chlamydia trachomatis infection. Sci. Signal. 3, ra21 (2010).
Fuerer, C. & Nusse, R. Lentiviral vectors to probe and manipulate the Wnt signaling pathway. PLoS One 5, e9370 (2010).

Patient-derived and mouse endo-ectocervical organoid generation, genetic manipulation and applications to model infection

推荐阅读