urine-based liquid biopsy for diagnostic of urogenital cancer

urine-based liquid biopsy for diagnostic of urogenital cancer

1. Urine- based Liquid Biopsy for Diagnosis /Screening of Urogenital Cancers Guorong Li, MD., Ph.D , HDR Dept of Urology /Plateau of Biology CHU Saint Etienne, France Mohammédia , November 2023

2. Urogenital or genitourinary cancers Immunological Implications and Molecular Diagnostics of Genitourinary Cancer. Editor: Moulay Mustapha Ennaji, Academic Press, 2023, Pages 5-18, ISBN 9780323854962, https://doi.org/10.1016/B978-0-323-85496-2.00032-4.

3. Tissue biopsy: Types of biopsy procedures used to diagnose cancer Needle biopsy (FNA and Core Biopsy) Skin biopsy Bone marrow biopsy Endoscopic biopsy Surgical biopsy

4. Liquid biopsy : recent and innovate method Lodewijk I, et al. Liquid Biopsy Biomarkers in Bladder Cancer: A Current Need for Patient Diagnosis and Monitoring. Int J Mol Sci . 2018 Aug 24;19(9):2514.

5. Tissue Biopsy vs Liquid Biopsy Michael Moore. WHAT ARE LIQUID BIOPSIES AND HOW ARE THEY USED IN THE TREATMENT OF CANCER? https://www.h-h-c.com/what-are-liquid-biopsies-and-how-are-they-used-in-the-treatment-of-cancer/

6. Urine-base liquid biopsy : urine exosomes Exosomes , nanoparticules. A recent subject promising for the discovery of biomarkers . A direct reflect of tumoral genetic and membrane markers.

7. Biogenesis and release of urine exosome

8. Diagnostic evolution : imaging + biopsy Malignant or benign ? Role of preoperative tissue biopsy . Diagnosis of Renal Cancer

9. Renal tissue biopsy Li G , et al. The use of MN/CA9 gene expression in identifying malignant solid renal tumors. European Urology, 49: 401-405, 2006 . Li G , et al. CA9 level in renal cyst fluid: a possible molecular diagnosis of malignant tumours . Histopathology 54: 880-884, 2009. Li G , et al. CA9 as a molecular marker for differential diagnosis of cystic renal tumors. Urologic Oncology 30: 463-468, 2012. Li G , et al. The level of serum CA9 is associated with postoperative recurrence in conventional renal cell cancer. Journal of Urology 180: 510-514, 2008. Li G, et al. CA9 gene expression in conventional renal cell carcinoma: a potential marker for prediction of early metastasis after nephrectomy. Clinical & Experimental Metastasis 24: 149-155, 2007. Li G , et al. MN/CA9 as a novel molecular marker for the detection of cancer. Expert Opinion on Medical Diagnostic 1: 91-97, 2007. Li G , et al, MN/CA9: a potential gene marker for detection of malignant cells in effusions. Biomarkers 12: 214-220, 2007.

10. Analysis of exosomes by TEM, FCM et RT- qPCR : Exosome markers: CD9, CD63 et CD81, Markers of renal clear cell cancer: CA9 et VEGFR2 . Li G , Mallouk N, Flandrin P, et al. Presence of Urinary Exosomes for Liquid Biopsy of Clear Cell Renal Cell Carcinoma : Protocol for a Pilot Feasibility Study . JMIR Res Protol , 2021 Ful 20;10(7);e24423, doi : 10,2196/24423, Isolation of exosomes

11. Mallouk N, Tan S, Scepaniack C, Garcin A, Pradat C, Lambert C, Li G . A specific technique of immunolabelling of urinary small extracellular vesicle biomarkers for the diagnostic of renal cancer. Microscopy ( Oxf ). 2023 Jan 27:dfad007. doi : 10.1093/ jmicro /dfad007. Epub ahead of print . PMID: 36715074 Li G , Mallouk N, Cornillon P. Urine-based liquid biopsy: a new avenue for the management of renal cell cancer. ExRNA 2022;4:23.

12. Diagnosis of Bladder Cancer: Urinary Cytology + Cystoscopy + Tissue Biopsy Imaging tests.  Imaging tests, such as computerized tomography (CT) urogram or retrograde pyelogram, allow the doctor to examine the structures of your urinary tract.

13. Wen J, Yang T, Mallouk N, Zhang Y, Li H, Lambert C,  Li G. Urinary Exosomal CA9 mRNA as a Novel Liquid Biopsy for Molecular Diagnosis of Bladder Cancer. Int J Nanomedicine . 2021 Jul 14;16:4805-4811. doi : 10.2147/IJN.S312322. eCollection 2021.

14. Diagnosis of Prostate Cancer PSA > 4 ng /ml or Abnormal DRE Usually 12 «  cores  »of tisues are taken using transrectal ultrasound (TRUS) guidance

15. HPV and Cervical Cancer Opportunity for prevention : Vaccination + Screening + Treatment 2030 goal: Eliminate cervical cancer: In 2020, the World Health Organization launched their cervical cancer elimination strategy. Introducing routine cervical cancer screening and treatment of cervical lesions is a key to achieving this goal.

16. Cervical Cancer: Colposcopy + Tissue Biopsy Necessity for new methods for diagnosis /screening Acceptable Less invasive Less expensive Fewer clinical visits Less need for clinical staff

17. Bergeron C, Orth G. La prévention du cancer du col utérin. Med Sci (Paris). 2023 May;39(5):423-428. French. doi : 10.1051/ medsci /2023057. Epub 2023 May 23. PMID: 37219346 Screening of Cervical Cancer

18. Global market of HPV test https://www.grandviewresearch.com/industry-analysis/hpv-testing-pap-test-market

19. HPV Tests: which standards? Poljak M, et al. Commercially available molecular tests for human papillomaviruses : a global overview . Clin Microbiol Infect. 2020:1144-1150.

20. Cervical cancer screening in Africa VIA (visual inspection with ascetic acid) or cytology used for cervical cancer screening. Moving from VIA or cytology to HPV testing. However, until now, HPV testing is not frequently available in African countries (performed at private facilities only against payment).

21. Human papillomavirus testing using existing nucleic acid testing platforms Joseph J,et al. Human papillomavirus testing using existing nucleic acid testing platforms to screen women for cervical cancer: implementation studies from five sub-Saharan African countries. BMJ Open. 2023 Jan 6;13(1):e065074.

22. HPV test in Africa : Challenges In terms of screening platform, the capacity to perform molecular tests has been significantly increased in most African countries to deal with the COVID-19 pandemic. Many platforms that are currently in use for HIV and TB testing can be used for HPV testing. The price of HPV tests can be reduced to significantly facilitate rapid transition from VIA-based to HPV DNA test based screening.

23. Self-sampled and clinician-sampled HPV test in Africa More than 95% of self-samples yielded HPV DNA results. The concordance in test results between self-collected samples and clinician-collected samples was reasonably high in most studies. Women of a wide age accepted self-sampling more easily. Self-collection of upper vaginal samples is likely to be the preferred approach over provider-collection. Nodjikouambaye ZA, et al. A systematic review of self- sampling for HPV testing in Africa . Int J Gynaecol Obstet . 2020 May;149(2):123-129. doi : 10.1002/ijgo.13112. Epub 2020 Feb 27. PMID: 32037532.

24. Urine HPV test: First void urine Poljak M, et al. Testing for Human Papillomaviruses in Urine, Blood, and Oral Specimens : an Update for the Laboratory . J Clin Microbiol . 2023 Jul 13:e0140322. doi : 10.1128/jcm.01403-22.

25. Kaoma, M., Olayemi , O., Mwaba , M.H. et al. Utilizing first void urine for high- risk HPV testing for cervical cancer screening in HIV-positive women in Katete , Zambia . BMC Women's Health 23, 62 (2023).

26. Cervical samples versus first void urine samples from India Purwar , Shashank , et al. Study to Determine Concordance between High-Risk Human Papilloma Virus DNA Detection in Self Collected First Voided Urine Samples and Health -Care Worker Collected Cervical Samples in a Subset of Women with Proven Histopathological Precancerous and Cancerous Lesions of the Cervix. Journal of Mid -life Health 14(1):p 8-14, Jan–Mar 2023. Conclusion: HPV DNA detection from the urine and cervical samples showed significant agreeability for the detection of precancerous and cancerous lesions of the cervix among women with abnormal histology results.

27. Kremer WW,, et al. The use of host cell DNA methylation analysis in the detection and management of women with advanced cervical intraepithelial neoplasia: a review. BJOG. 2021 Feb;128(3):504-514. doi : 10.1111/1471-0528.16395, Methylation analysis for cervical cancer

28. Molecular test of triage for positive HPV Pun , P.B., Liao, YP., Su, PH. et al. Triage of high- risk human papillomavirus-positive women by methylated POU4F3 . Clin Epigenet 7, 85 (2015). https://doi.org/10.1186/s13148-015-0122-0 Tian Y., et al Utility of gene methylation analysis , cytological examination , and HPV-16/18 genotyping in triage of high- risk human papilloma virus-positive women . Oncotarget . 2017; 8: 62274-62285.

29. Methylation analysis in urine samples van den Helder R, et al. HPV and DNA Methylation Testing in Urine for Cervical Intraepithelial Neoplasia and Cervical Cancer Detection . Clin Cancer Res . 2022 May 13;28(10):2061-2068.

30. Urine HPV test: Preliminary results from Mohammedia and Saint–Etienne n° Cervical sample Urine sample 1 + - 2 + + 3 + + 4 + - 5 + + 6 + + 7 + + 8 + + 9 + - 10 - - 11 - - 12 - - 13 - - 14 - - 15 - - 16 - - 17 - - 18 - - 19 - - 20 - - n° Cervical sample Urine sample 1 58 58 2 18, 51 18, 51 3 - 58 4 31, 51, 59 31, 51, 59 5 31, 45, 68 31, 45 6 16 16 7 52 - 8 - 66 9 51, 53 53, 54 10 16 16 11 16 16 12 31, 54, 58, 68, 73 31, 54, 58, 68, 73 13 - 51 14 31 31 15 - - 16 16 51, 33 17 16 16 18 16, 56 16, 56 19 31, 51 - 20 - - Mohammedia: DNA extraction Invitrogen + nested PCR (HPV L1 gene : a broad spectrum of HPV types). Saint Etienne: Automate Nimbus extraction of DNA and Anyplex ™ II HPV HR Detection PCR of HPV DNA.

31. Urine-based detection of HPV for cervical cancer screening Li G, Lamsisi M, Chenafi S, Moniod L, Benlghazi A, Ennaji MM, Chauleur C, Bourlet T. Urine- based detection of HPV for cervical cancer screening: Time for standardized tests. J Med Virol . 2023 Apr;95(4):e28737. doi : 10.1002/jmv.28737. PMID: 37185853. Li G, Moniod L, Chenafi S, Lamsisi M, Ennaji MM, Bourlet T, Chauleur C. Detection of HPV in urine for cervical cancer screening: Feasibilty of an assay system. J Gynecol Obstet Hum Reprod . 2023 Jul 12;52(8):102631. doi : 10.1016/j.jogoh.2023.102631. Epub ahead of print . PMID: 37451412. Lamsisi M , Li G , Chauleur C, Ennaji M, Bourlet T. The potential of urine for human papillomavirus- related cervical cancer prevention . FUTURE VIROLOGY 17 (7), 2022. Published Online:29 Apr 2022 https://doi.org/10.2217/fvl-2021-0246 1: How to collect urine: Pre- analysis (Collection, Preservation , Transportation)? 2: How to analyze : international standards or laboratory-based PCR (HPV genotype or HPV16/18 or HPV +/-? 3: Which role of urine HPV detection in Africa : efficacity versus cost ?

32. Urine-based liquid biopsy for diagnosis/screening of urogenital cancer is promising. Detection of HPV in urine can be used for cervical cancer screening. Implementing standardized self-sampling techniques and laboratory-based HPV assays is paramount for cervical cancer screening. Conclusions

33. Thank you for your attention Attention: These are not the urine samples