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Nano-bioconjugate epigenetic modulator carriers as a therapeutic strategy in breast cancer.
Brazil (IFMSA-Brazil) - Universidade Federal do Parana (UFPR), Curitiba
Profª Giseli Klassen, PhD
Profª Giseli Klassen, PhD
Type of Research Project
- Basic science
What is the background of the project?
There are few therapeutic options for patients with metastatic or invasive breast cancer, and even those affected by less aggressive tumors require prolonged treatment. Conventional chemotherapy treatments for breast cancer still lack specificity and consequently cause irreversible damage to both tumor tissue and healthy cells. The aim of this project is to develop theranostic nanoparticles containing selective nano-bioconjugates to human epidermal growth factor receptor 2 (HER2) - a biomarker overexpressed breast tumor cells - and to analyze the effect of decitabine demethylating drug in vitro and in vivo. The project will analyze the selective vectoring ability for invasive tumor cells and the antitumor effect and selectivity of the nano-bioconjugates. The proposed nanotheranostic agent has the potential for selective accumulation in invasive breast tumors, allowing (1) cancer cell detection through real-time fluorescence extracorporeal imaging and (2) promoting decitabine release in order to restore the expression of tumor suppressor genes silenced by epigenetic mechanisms. The generation of the proposed system would bring numerous benefits, such as greater antitumor activity, lower side effects, reduction of the total required dose of chemotherapy and even decreasing the final cost of treatment.
What is the aim of the project?
The aim of this project is to develop theranostic nanoparticles containing selective nano-bioconjugates to human epidermal growth factor receptor 2 (HER2) - a biomarker overexpressed breast tumor cells - and to analyze the effect of decitabine drug in vitro and in vivo through expression of the disintegrin and metalloproteinase domain-containing protein 33 (ADAM33) gene.
What techniques and methods are used?
Techniques and methods used: F1. Synthesize and characterize PLA-PEG-antiHER2 polymer by proton nuclear magnetic resonance (1H NMR), gel permeation chromatography (GPC) and Fourier-transform infrared spectroscopy (FTIR). 2. Formulate nanoparticles with different proportions between PLA-PEG and PLA-PEG-antiHER2 containing Nile red and characterize them by dynamic light scattering, zeta potential, transmission electron microscopy and confocal microscopy. 3. Analyze cell viability, selectivity and binding affinity of nanoparticles in in vitro breast tumor lines by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor (MTT) Quartz Crystal Microbalance with Dissipation monitoring (QCM-D), flow cytometry and Stochastic Optical Reconstruction Microscopy (STORM) confocal microscopy assays. 4. Incorporate the drug decitabine into the nanoparticle formulation and analyze the release profile, stability and effect on breast tumor lines in vitro. 5. Analyze the effect of incubating decitabine-loaded nanoparticles in vitro with ADAM33 expression and viability in breast tumor lines. 6. Analyze effect of decitabine-loaded nanoparticles in vivo by monitoring tumor remission, selectivity and extracorporeal fluorescence imaging.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student is expected to cultivate breast cancer cell lines in bottles,for example, the Michigan Cancer Foundation-7 (MCF-7) or Medical Doctor Anderson - Metastasis Breast Cancer 231 (MDA MB 231) cell lines, which involves changing the medium, expanding the "cell colonies", freezing and unfreezing cells. The student is also expect to isolate deoxyribonucleic acid and ribonucleic acid from the same cells, as well as gene expression evaluation through techniques such as reverse transcription followed by real time/quantitative polymerase chain reaction (RT-qPCR), Western Blotting, agarose gel electrophoresis. Other tasks will depend on which stage of the project the student will work with, as already described in the "techniques and methods" section of this form.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The tutor or the local students will personally teach the incoming the theory and techniques required at any specific moment.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a presentation - The student will prepare a scientific report - The student will prepare an abstract - The student’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Subjects passed: biochemistry, molecular cell biology, basic concepts of chemistry,
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system - Students in biomedical fields
- E.A. Ramos; A.A. Camargo; K. Braun; R. Slowik; I.J. Cavalli; E.M. Ribeiro; O. Pedrosa Fde; E.M. de Souza; F.F. Costa; G. Klassen; Simultaneous CXCL12 and ESR1 CpG island hypermethylation correlates with poor prognosis in sporadic breast cancer; BMC Cancer; 10 (2010) 23.
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