Принципы и подходы к созданию флуоресцентных гидрогелей для диагностики онкологических заболеваний

Введение. Ранняя чувствительная и высокоспецифичная диагностика является одной из важных составляющих успешной терапии онкологических заболеваний. Флуоресцентные гидрогели позволяют создавать универсальные биосенсоры благодаря повышенной емкости связывания биологических распознающих и репортерных молекул, возможности проведения высокочувствительной флуоресцентной детекции, а также гибкости комбинирования их структурных и функциональных элементов.

Цель исследования – рассмотреть принципы дизайна и методические подходы к созданию биосенсоров на основе флуоресцентных гидрогелей для детекции онкомаркеров, а также обобщить и систематизировать данные по применяемым в них принципам детекции и генерации детектируемого сигнала.

Результаты. Флуоресцентные гидрогели являются примером трехмерных сенсорных платформ – структур, объединяющих репортерную флуоресцентную функцию с биологическими распознающими молекулами, которые позволяют сохранить уникальные оптические свойства флуоресцентных нанокристаллов на макроуровне. Пористая структура гидрогелей позволяет увеличить активную площадь поверхности биосенсоров для трехмерной иммобилизации флуоресцентных меток и биологических распознающих молекул, при этом сохраняя структуру последних для специфического связывания детектируемых молекул образца, что обеспечивает чувствительность, превосходящую традиционные методы иммуноферментного и иммунохроматографического анализа. В качестве биологических распознающих молекул могут выступать не только традиционно применяемые антитела, но и ферменты и гликопротеины, аптамеры и олигонуклеотиды, а также полимеры, полученные методом молекулярного импринтинга, что расширяет круг специфически детектируемых аналитов.

Заключение. В обзоре представлены примеры биосенсоров на основе флуоресцентных гидрогелей для детекции маркеров онкологических заболеваний, изложены подходы к получению этих гелей, иммобилизации биологических распознающих молекул и принципы генерации оптического детектируемого сигнала. Показаны основные преимущества флуоресцентных гидрогелевых биосенсоров перед классическими тестами, применяемыми в области экспресс-диагностики онкологических заболеваний.

1. Hawkes N. Cancer survival data emphasise importance of early diagnosis. BMJ 2019;364:1408. DOI: 10.1136/bmj.l408

2. Crosby D., Bhatia S., Brindle K.M. et al. Early detection of cancer. Science 2022;375(6586):eaay9040. DOI: 10.1126/science.aay9040

3. Pulumati A., Pulumati A., Dwarakanath B.S. et al. Technological advancements in cancer diagnostics: Improvements and limitations. Cancer Rep (Hoboken) 2023;6(2):e1764. DOI: 10.1002/cnr2.1764

4. Sukhanova A., Ramos-Gomes F., Chames P. et al. Multiphoton deep-tissue imaging of micrometastases and disseminated cancer cells using conjugates of quantum dots and single-domain antibodies. Methods Mol Biol 2021;2350:105–23. DOI: 10.1007/978-1-0716-1593-5_8

5. Lutz A.M., Willmann J.K., Cochran F.V. et al. Cancer screening: a mathematical model relating secreted blood biomarker levels to tumor sizes. PLoS medicine 2008;5(8):e170. DOI: 10.1371/journal.pmed.0050170

6. You P.Y., Li F.C., Liu M.H., Chan Y.H. Colorimetric and fluorescent dual-mode immunoassay based on plasmon-enhanced fluorescence of polymer dots for detection of PSA in whole blood. ACS Appl Mater Interfaces 2019;11(10):9841–9. DOI: 10.1021/acsami.9b00204

7. Li Y., Huang Z.-Z., Weng Y., Tan H. Pyrophosphate ion-responsive alginate hydrogel as an effective fluorescent sensing platform for alkaline phosphatase detection. Chem Commun (Camb) 2019;55(76):11450–3. DOI: 10.1039/C9CC05223B

8. Guglielmi M., Martucci A. Semiconductor quantum dot-doped sol–gel materials. In: Martucci A., Santos L., Estefanía Rojas Hernández R., Almeida R., eds. Sol–gel derived optical and photonic materials. Cambridge: Woodhead Publishing, 2020. P. 209–226.

9. Richter A., Paschew G., Klatt S. et al. Review on hydrogel-based pH sensors and microsensors. Sensors (Basel) 2008;8(1):561–81. DOI: 10.3390/s8010561

10. Zhang Z., He C., Chen X. Hydrogels based on pH-responsive reversible carbon–nitrogen double-bond linkages for biomedical applications. Mat Chem Front 2018;2:1765–78. DOI: 10.1039/C8QM00317C

11. Hashim H., Maruyama H., Akita Y. et al. Hydrogel fluorescence microsensor with fluorescence recovery for prolonged stable temperature measurements. Sensors (Basel) 2019;19(23):5247. DOI: 10.3390/s19235247

12. Jia Z., Sukker I., Müller M., Schönherr H. Selective discrimination of key enzymes of pathogenic and nonpathogenic bacteria on autonomously reporting shape-encoded hydrogel patterns. ACS Appl Mater Interfaces 2018;10(6):5175–84. DOI: 10.1021/acsami.7b15147

13. Liang Z., Zhang J., Wu C. et al. Flexible and self-healing electrochemical hydrogel sensor with high efficiency toward glucose monitoring. Biosens Bioelectron 2020;155:112105. DOI: 10.1016/j.bios.2020.112105

14. Chen Z., Chen Y., Hedenqvist M.S. et al. Multifunctional conductive hydrogels and their applications as smart wearable devices. J Mater Chem B 2021;9(11):2561–83. DOI: 10.1039/D0TB02929G

15. Larsson A., Ekblad T., Andersson O., Liedberg B. Photografted poly(ethylene glycol) matrix for affinity interaction studies. Biomacromolecules 2007;8(1):287–95. DOI: 10.1021/bm060685g

16. Jung I.Y., Kim J.S., Choi B.R. et al. Hydrogel based biosensors for in vitro diagnostics of biochemicals, proteins, and genes. Adv Healthc Mater 2017;6(12):1601475. DOI: 10.1002/adhm.201601475

17. Gao Y., Wolf L.K., Georgiadis R.M. Secondary structure effects on DNA hybridization kinetics: A solution versus surface comparison. Nucleic Acids Res 2006;34(11):3370–7. DOI: 10.1093/nar/gkl422

18. Welch N.G., Scoble J.A., Muir B.W., Pigram P.J. Orientation and characterization of immobilized antibodies for improved immunoassays (Review). Biointerphases 2017;12(2):02D301. DOI: 10.1116/1.4978435

19. Feng B., Huang S., Ge F. et al. 3D antibody immobilization on a planar matrix surface. Biosens Bioelectron 2011;28(1):91–6. DOI: 10.1016/j.bios.2011.07.003

20. Su X., Hao D., Xu X. et al. Hydrophilic/hydrophobic heterogeneity anti-biofouling hydrogels with well-regulated rehydration. ACS Appl Mater Interfaces 2020;12(22):25316–23. DOI: 10.1021/acsami.0c05406

21. Missirlis D., Baños M., Lussier F., Spatz J.P. Facile and versatile method for micropatterning poly(acrylamide) hydrogels using photocleavable comonomers. ACS Appl Mater Interfaces 2022;14(3):3643–52. DOI: 10.1021/acsami.1c17901

22. Xia Y., Xue B., Qin M. et al. Printable fluorescent hydrogels based on Self-assembling peptides. Sci Rep 2017;7(1):9691. DOI: 10.1038/s41598-017-10162-y

23. Kar T., Patra N. Pyrene-based fluorescent supramolecular hydrogel: Scaffold for nanoparticle synthesis. J Phys Org Chem 2019;33:e4026. DOI: 10.1002/poc.4026

24. Wu Y., Jin X., Ashrafzadeh Afshar E. et al. Simple turn-off fluorescence sensor for determination of raloxifene using gold nanoparticles stabilized by chitosan hydrogel. Chemosphere 2022;305:135392. DOI: 10.1016/j.chemosphere.2022.135392

25. Liu C., Li Q., Wang H. et al. Quantum dots-loaded self-healing gels for versatile fluorescent assembly. Nanomaterials (Basel) 2022;12(3):452. DOI: 10.3390/nano12030452

26. Pisanic T.R. 2nd, Zhang Y., Wang T.H. Quantum dots in diagnostics and detection: principles and paradigms. Analyst 2014;139(12):2968–81. DOI: 10.1039/c4an00294f

27. Linkov P.A., Vokhmintcev K.V., Samokhvalov P.S., Nabiev I.R. Ultrasmall quantum dots for fluorescent bioimaging in vivo and in vitro. Opt Spectrosc 2017;122(1):8–11. DOI: 10.1134/S0030400X17010143

28. Kandi D., Mansingh S., Behera A., ParidaK. Calculation of relative fluorescence quantum yield and Urbach energy of colloidal CdS QDs in various easily accessible solvents. J Lumin 2021;231:117792. DOI: 10.1016/j.jlumin.2020.117792

29. Neo D.C.J., Goh W.P., Lau H.H., Shanmugam J. CuInS2 quantum dots with thick ZnSexS1-x shells for a luminescent solar concentrator. ACS Appl Nano Mater 2020;3:6489–96. DOI: 10.1021/acsanm.0c00958

30. Dey S.C., Nath S.S. Size-dependent fluorescence in CdSe quantum dots. Emer Mat Res 2012;1(3):117–20. DOI: 10.1680/emr.11.00004

31. Montón H., Nogués C., Rossinyol E. et al. QDs versus Alexa: reality of promising tools for immunocytochemistry. J Nano-biotechnology 2009;7:4. DOI: 10.1186/1477-3155-7-4

32. Benson K., Ghimire A., Pattammattel A., Kumar C.V. Protein biophosphors: Biodegradable, multifunctional, protein-based hydrogel for white emission, sensing, and pH detection. Adv Funct Mater 2017;27:1702955. DOI: 10.1002/adfm.201702955

33. Li C.Y., Zheng S.Y., Du C. et al. Carbon dot/poly(methylacrylic acid) nanocomposite hydrogels with high toughness and strong fluorescence. ACS Appl Polym Mater 2020;2:1043–52. DOI: 10.1021/acsapm.9b00971

34. Xu J., Zhang Y., Zhu W. et al. Synthesis of Polymeric nanocomposite hydrogels containing the pendant ZnS nanoparticles: Approach to higher refractive index optical polymeric nanocomposites. Macromol 2018;51(Is.7):2672–81. DOI: 10.1021/acs.macromol.7b02315

35. Yang T., Li Q., Wen W. et al. Cadmium sulfide quantum dots/ poly(acrylic acid-co-acrylic amide) composite hydrogel synthesized by gamma irradiation. Rad Phys Chem 2018;145:130–4. DOI: 10.1016/j.radphyschem.2017.10.012

36. Zhang H., Wang X., Liao Q. et al. Embedding perovskite nanocrystals into a polymer matrix for tunable luminescence probes in cell imaging. Advan Func Mat 2017;27(7):1604382. DOI: 10.1002/adfm.201604382

37. Gaponik N., Wolf A., Marx R. et al. Three-dimensional self-assembly of thiol-capped CdTe nanocrystals: Gels and aerogels as building blocks for nanotechnology. Advan Mat 2008;20(Is.22):4257–62. DOI: 10.1002/adma.200702986

38. Hörner M., Becker J., Bohnert R. et al. A photoreceptor-based hydrogel with red light-responsive reversible sol-gel transition as transient cellular matrix. Adv Mat Tech n/a:2300195. DOI: 10.1002/admt.202300195

39. Bhattacharya S., Nandi S., Jelinek R. Carbon-dot–hydrogel for enzyme-mediated bacterial detection. RSC Advances 2017;7:588–94. DOI: 10.1039/C6RA25148J

40. Lee T., Teng T.Z.J., Shelat V.G. Carbohydrate antigen 19-9 – tumor marker: Past, present, and future. World J Gastrointest Surg 2020;12(12):468–90. DOI: 10.4240/wjgs.v12.i12.468

41. Piloto A.M.L., Ribeiro D.S.M., Rodrigues S.S.M. et al. Cellulose-based hydrogel on quantum dots with molecularly imprinted polymers for the detection of CA19-9 protein cancer biomarker. Mikrochim Acta 2022;189(4):134. DOI: 10.1007/s00604-022-05230-8

42. Ahmadi-Sangachin E., Mohammadnejad J., Hosseini M. Fluorescence Self-assembled DNA hydrogel for the determination of prostate specific antigen by aggregation induced emission. Spectrochim Acta A Mol Biomol Spectrosc 2023:303:123234. DOI: 10.2139/ssrn.4313053

43. Bautista-Sánchez D., Arriaga-Canon C., Pedroza-Torres A. et al. The promising role of miR-21 as a cancer biomarker and its importance in RNA-based therapeutics. Mol Ther Nucleic Acids 2020;20:409–20. DOI: 10.1016/j.omtn.2020.03.003

44. Mohammadi S., Mohammadi S., Salimi A. A 3D hydrogel based on chitosan and carbon dots for sensitive fluorescence detection of microRNA-21 in breast cancer cells. Talanta 2021;224:121895. DOI: 10.1016/j.talanta.2020.121895

45. Gao Y., Feng B., Han S. et al. The roles of microRNA-141 in human cancers: From diagnosis to treatment. Cell Physiol Biochem 2016;38(2):427–48. DOI: 10.1159/000438641

46. Li C., Li H., Ge J., Jie G. Versatile fluorescence detection of microRNA based on novel DNA hydrogel-amplified signal probes coupled with DNA walker amplification. Chem Commun (Camb) 2019;55(27):3919–22. DOI: 10.1039/C9CC00565J

47. Gong X., Zhou W., Chai Y. et al. MicroRNA-induced cascaded and catalytic self-assembly of DNA nanostructures for enzyme-free and sensitive fluorescence detection of microRNA from tumor cells. Chem Commun (Camb) 2016;52(12):2501–4. DOI: 10.1039/C5CC08861E

48. Zhang G., Zhou S., Yan G. et al. Quantum dot-crosslinked light-guiding hydrogels for sensing folate receptor-overexpressed cancer cells. Sens Actuators B Chem 2021;349:130815. DOI: 10.1016/j.snb.2021.130815

49. Bolli A., Galluzzo P., Ascenzi P. et al. Laccase treatment impairs bisphenol A-induced cancer cell proliferation affecting estrogen receptor alpha-dependent rapid signals. IUBMB life 2008;60(12):843–52. DOI: 10.1002/iub.130

50. Ruiz-Palomero C., Benítez-Martínez S., Soriano M.L., Valcárcel M. Fluorescent nanocellulosic hydrogels based on graphene quantum dots for sensing laccase. Anal Chim Acta 2017;974:93–9. DOI: 10.1016/j.aca.2017.04.018

51. Tse R.T.-H., Wong C.Y.-P., Chiu P.K.-F., Ng C.F. The potential role of spermine and its acetylated derivative in human malignancies. Int J Mol Sci 2022;23(3):1258. DOI: 10.3390/ijms23031258

52. Nair R.R., Debnath S., Das S. et al. A highly selective turn-on biosensor for measuring spermine/spermidine in human urine and blood. ACS Appl Bio Mater 2019;2(6):2374–87. DOI: 10.1021/acsabm.9b00084

53. Traverso N., Ricciarelli R., Nitti M. et al. Role of glutathione in cancer progression and chemoresistance. Oxid Med Cell Longev 2013;2013:972913. DOI: 10.1155/2013/972913

54. Wu R., Ge H., Liu C. et al. A novel thermometer-type hydrogel senor for glutathione detection. Talanta 2019;196:191–6. DOI: 10.1016/j.talanta.2018.12.020

55. Grant C.E., Flis A.L., Ryan B.M. Understanding the role of dopamine in cancer: Past, present and future. Carcinogenesis 2022;43(6):517–27. DOI: 10.1093/carcin/bgac045

56. Yuan J., Wen D., Gaponik N., Eychmüller A. Enzyme-encapsulating quantum dot hydrogels and xerogels as biosensors: Multifunctional platforms for both biocatalysis and fluorescent probing. Angew Chem Int Ed Engl 2013;52(3):976–9. DOI: 10.1002/anie.201205791

57. Fini M.A., Elias A., Johnson R.J., Wright R.M. Contribution of uric acid to cancer risk, recurrence, and mortality. Clin Transl Med 2012;1(1):16. DOI: 10.1186/2001-1326-1-16

58. Azmi N.E., Rashid A.H.A., Abdullah J. et al. Fluorescence biosensor based on encapsulated quantum dots/enzymes/sol-gel for non-invasive detection of uric acid. J Luminescence 2018;202:309–15. DOI: 10.1016/j.jlumin.2018.05.075

59. Wang M., Zhu J., Lubman D.M., Gao C. Aberrant glycosylation and cancer biomarker discovery: a promising and thorny journey. Clin Chem Lab Med 2019;57(4):407–16. DOI: 10.1515/cclm-2018-0379

60. Koshi Y., Nakata E., Yamane H., Hamachi I. A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates. J Am Chem Soc 2006;128(32):10413–22. DOI: 10.1021/ja0613963