The development of innovative technologies increasing diagnosis and treatment efficiency of the cervix background and precancerous diseases associated with HPV

Individual registration number ВR18574160

Еxecution period: 2023 -2024

Source of funding - Ministry of Science and Higher Education of the Republic of Kazakhstan

The head of the research group: Shanazarov Nasrulla Abdullayevich

The project aims. Development of innovative technologies that increase the efficiency of diagnosis and treatment of background and precancerous diseases of the cervix associated with HPV.

Project tasks.

1. Evaluation of the effectiveness and safety of the applied methods of treatment of background and precancerous diseases of the cervix associated with HPV.
2. Development of an improved, effective and safe method for the treatment of background and precancerous diseases of the cervix associated with HPV, using laser technologies with photosensitization (then - PS). 
3. Development of a method of fluorescent diagnostics using PS for background and precancerous diseases of the cervix associated with HPV. 
4. Development of a method for assessing the "burnout" of FS during local photodynamic therapy (then- PDT). 
5. Development and implementation of an optimal PDT program for complete eradication of HPV. 

Expected results.

• For the first time, immunological and molecular genetic data will be obtained reflecting the causes of HPV persistence and the duration of its elimination when using complex PDT technology
• For the first time in the Republic of Kazakhstan, an innovative complex laser technology of light therapy with photosensitization will be developed and used for the treatment of HPV-associated precancerous diseases of the cervix, taking into account the endotype.
• 3 articles and (or) reviews will be published in peer-reviewed scientific publications in the scientific direction of the program, included in the 1, 2  and (or) 3 quartile by the impact factor in the database Web of Science data and (or) having a CiteScore percentile in the Scopus database of at least 50, 3 articles in peer-reviewed foreign or domestic publications recommended by Committee for Quality Assurance in the Sphere of Education and Science of the Ministry of Education and Science of the Republic of Kazakhstan, abstracts and 2 reports at international scientific and practical conferences, a patent of the Kazakhstan Patent Bureau, methodological recommendations, suggestions for periodic clinical protocols, monograph.

Final Results According to the 2023–2024 Calendar Plan

In accordance with the set objectives, the following types and stages of research work were completed:

1. Genetic Analysis and Formation of a Candidate Gene Panel

Based on international studies, a panel of candidate genes was compiled for whole-exome sequencing (WES) aimed at identifying genetic factors potentially involved in the pathogenesis of cervical cancer. The panel includes genes associated with:

• Immune response and inflammation: HLA-B, MICA, IFNG, TNF, TGFβ1, IL10, IL6, CTLA4, CD83
• Apoptosis and caspase pathways: TP53, CASP8, FAS, FASLG, BAX, BID, MCL1
• Cell cycle regulation and DNA repair: PIK3CA, PTEN, ATM, BRCA1/2, CHEK2, FANCA, FANCM, RAD51B, PALB2
• Oncogenes and transcription factors: MYC, CCND1, E2F1, ERBB2/3/4, EGFR, NRAS, KRAS
• Toll-like receptors and signaling cascades: TLR2, TLR3, TLR4, TLR9, NFE2L2, KEAP1, MAPK1

Bioinformatic analysis of 192 DNA samples identified variant differences in these priority genes compared to reference genomic sequences.

2. Application and Evaluation of Photodynamic Therapy (PDT)

An optimized, comprehensive PDT program was implemented for 150 patients diagnosed with HPV-associated cervical dysplasia. The protocol included:

• Intravenous administration of the photosensitizer “Photolon” (1.2 mg/kg);
• Intravenous laser blood irradiation (wavelength 662 nm, power 0.1 W, duration 30 minutes);
• Local irradiation of the cervix under fluorescence-guided control using flexible and surface light guides (500–1200 mW, duration 6–21 minutes depending on location).

A method of fluorescence diagnostics (FD) was developed and used at multiple time points—before PDT, during follow-up, and after treatment. A novel “photosensitizer burnout” assessment technique was introduced, allowing for better differentiation between affected and healthy tissues by enhancing red/blue contrast in fluorescence imaging.

3. Immunological and Clinical Evaluation of PDT Effectiveness

A comprehensive evaluation was conducted in 200 patients (150 in the treatment group, 50 in the control group), which included:

• Immunological profiling (CD3, CD4, CD8, CD16, CD56, CD19, IL-1, IL-6, TNF-α) before PDT, and at 5 days, 3 months, and 12 months post-treatment;
• Clinical assessment before therapy and 3 hours after;
• HPV and urogenital infection screening using PCR (600 tests);
• Collection of phenotypic, anthropometric, and molecular-genetic data.

All collected data were entered into an Excel database, which is being continuously updated.

4. Scientific and Patent Output

Patents:

1. Method for treating background and precancerous cervical diseases — National Institute of Intellectual Property (Kazakhstan), IPC A61N 5/06; A61P 35/00

2. Method for treating background and precancerous cervical diseases — Federal Service for Intellectual Property (Russia), IPC A61K 31/409

Key Scientific Publications:

1. Shanazarov et al. Photodynamic therapy of cervical cancer // Ther Adv Chronic Dis, 2024 — Percentile 79

2. Shanazarov et al. Effectiveness and safety of photodynamic therapy in CIN // Photodiagnosis Photodyn Ther, 2024 — Percentile 86

3. Aitkaliyev et al. Challenges of false positive cytological results... // Science and Healthcare, 2023

4. Smailova et al. Fluorescence diagnostics in cervical dysplasia // Biomedical Photonics, 2023

Methodological Guidelines:

• Published in Russian and Kazakh: Photodynamic therapy of HPV-associated background and precancerous cervical diseases
• Approved for publication by the National Scientific Center for Health Development, Ministry of Health of the Republic of Kazakhstan (Protocol No. 478, Sept 16, 2024)
• A chapter titled Photodynamic Therapy of Precancerous Diseases of Female Reproductive Organs published in the monograph Photodynamic Theranostics

5. Participation in Scientific Conferences

Congress and Conference Presentations:

• XII and XIII International Congresses on Photodynamic Therapy and Diagnostics (Moscow, 2023–2024)
• XIV Congress of Oncologists and Radiologists of the CIS and Eurasia (Dushanbe, 2024)
• Topics presented included:
• Priority directions of PDT in oncology
• Comprehensive treatment approach for cervical precancer within the national health development strategy
• Comparative analysis of legal frameworks for PDT implementation
• Fluorescence diagnostics for identifying multicentric foci of cervical dysplasia
• PDT effects on Toll-like receptors (TLR2, 3, 4, 8) in HPV-associated cervical dysplasia

Conference Abstracts Published in “Eurasian Oncology Journal”:

1. First experience of PDT in reproductive-age women with HPV-associated dysplasia

2. Effectiveness of PDT in HPV-related squamous intraepithelial cervical lesions

3. Candidate genes in patients with HPV-associated cervical dysplasia

4. PDT effects on mucosal immunity markers in precancer and cervical cancer treatment

Conclusion

The research goals were successfully met:

• A comprehensive PDT protocol was developed and implemented for HPV-associated cervical lesions;
• A structured database was created, incorporating clinical, immunological, and genetic data for 200 patients;
• Patents were obtained and numerous peer-reviewed publications produced;
• Fluorescence diagnostics and “photosensitizer burnout” assessment methods were introduced and optimized;
• The study gained international recognition through publications and conference presentations.

Contact details: Shanazarov Nasrulla Abdullayevich
+ 7 (7172) 70-80-90, ext. nom. 8081