Implicacions mol·leculars, biològiques i clíniques del sistema hidroxiolita en càncer de mama
- Autores: Anna Maria Rodriguez Rivero
- Directores de la Tesis: Pau Turon Dols (dir. tes.), Joan Francesc Julian Ibañez (codir. tes.)
- Lectura: En la Universitat Autònoma de Barcelona ( España ) en 2021
- Idioma: catalán
- Tribunal Calificador de la Tesis: Salvador Navarro Soto (presid.), Susana Puig Sardá (secret.), Manel Esteller Badosa (voc.)
- Programa de doctorado: Programa de Doctorado en Cirugía y Ciencias Morfológicas por la Universidad Autónoma de Barcelona
- Materias:
- Enlaces
- Tesis en acceso abierto en: TDX
- Resumen
Breast cancer is the most frequent cancer among women with two million new cases diagnosed in 2018. The understanding of its origin and progression through new approaches is of outmost interest. In this thesis, we contribute with a new line of research based on natural biomineralization of nucleic acids, transferring knowledge from materials science to clinical research. We focus on the molecular content of breast cancer calcifications, their role as natural non-viral vectors of transfection (NVT) and their expected relationship with breast cancer multifocality.
The feasibility of the nucleic acid biomineralization has recently been demonstrated by our group using experimental and computational methods with the ultimate intention to explore the relationship among calcifications found in tumor tissue, the process of DNA biomineralization and its biological function. We demonstrate in Revilla-López et al. (Langmuir. 2019; 35(36): 11912-11922 https://doi.org/10.1021/acs.langmuir.9b01566), by means of Molecular Dynamics simulation, that adsorption of nucleic acids in both hydroxyapatite (HAp) and calcium oxalate (CaOx) is feasible. However, only HAp can encapsulate those biopolymers. Such a hybrid structure composed of HAp and nucleic acids, we termed as hydroxyolite (HOLi), is identical to an inorganic NVT which is able to introduce the nucleic acids carried by the mineral into a target cell. We prove that DNA biomineralization, independently of its degree of methylation, is favored on HAp compared to CaOx. Moreover, we study the influence of Mg2+ ions and we unveil that Mg2+ facilitates DNA encapsulation in HAp but disables it in CaOx.
Secondly, we explore the existence of hydroxyolites in human breast cancer tissue freshly excised by using Raman spectroscopy. Through new advanced Raman imaging algorithm in combination with advance multivariate analysis (Marro et al., Cancers. 2021; 13(11):2658 https://doi.org/10.3390/cancers13112658) we confirm that HAp microcalcifications contain DNA adsorbed and encapsulated demonstrating the hypothesis that living tissue creates hybrid structures equal to a NVT.
Thirdly, we investigate if natural calcifications (i.e. amorphous calcium phosphate (ACP), HAp and CaOx) can act as NVT (Rodríguez-Rivero et al., 2021 – ready for submission). We conclude, using a known transfection model (A549 human lung epithelial cells), that all of them are able to transfect but transfection efficiency of ACP, a precursor of HAp, is much higher than HAp and CaOx. We study the effect of supplying Ca2+ and Mg2+, demonstrating that Ca2+ enhances the transfection of all of them and even makes feasible the transfection of naked DNA. On the other hand, we confirm that such NVT can migrate to a certain distance (up to 50 mm) before transfecting, simulating the clinical scenario of multifocality.
The study of DNA biomineralization in tumor tissue, the migration of HAp-DNA particles that protect DNA from enzymatic degradation and the capacity of transfection of natural NVT open new avenues in cancer research. The findings of this thesis contribute to the understanding of the clinical scenario related to breast cancer multifocality.
