NUCEL GROUP OF CELLULAR AND MOLECULAR THERAPY

Environmental control technician at the Federal Institute of Education, Science, and Technology of Rio Grande do Norte (2008). Graduated in biological sciences (licentiate and bachelor’s degree) from the Federal University of Rio Grande do Norte – UFRN (2011). Master’s degree in biochemistry and molecular biology from UFNR (2014). Currently a doctoral student in biochemistry at the Institute of Chemistry Postgraduate Program at the University of São Paulo in partnership with the Center of Cellular and Molecular Therapy (NUCEL). Has experience in embryology and human anatomy working on the morphophysiological characterization of umbilical cords in dogs and histology of embryonic systems. Has experience in biochemistry, and cellular and molecular biology working on purification/kinetic characterization of enzymes, metagenomics of extreme environments, and functional identification/characterization of molecular markers (overexpression and knockout via CRISPR/Cas9 gene-editing system) in human breast cancer cell lines.

Breast cancer is the most common type of cancer detected in women worldwide [1], being considered a serious global public health problem, whose early and correct diagnosis is still the main way to prevent disease progression. The use of molecules as tumor markers has helped clinicians in predictive diagnosis, prognostic assessment, and even in the treatment of breast cancer, being essential in reducing the high mortality rates caused by the spread of the disease to other regions of the body, especially the bones. However, only a small group of tumor markers are entirely reliable, stimulating the search for new targets, and their functional characterization, as a way to better understand the biology and molecular bases of this disease. 

Based on the analysis of gene expression and gain and loss of function experiments, our research group has been developing projects that aim to discover the relationship of proteincoding and non-protein-coding genes with the maintenance of normal cells, and with the progression of the tumor phenotype, thus helping in the development of new clinical and therapeutic protocols. In one of the group’s lines of research, focused on protein-coding genes, we describe the protein markers CD90 and CD14 as being differentially expressed in human breast cancer cell lines [2], which may be related to the degree of malignancy of the disease, therefore constituting interesting targets to be studied. Functional characterization experiments for the CD90 marker have already been carried out [3], while those for the CD14 marker are ongoing.

Nevertheless, studies that relate the expression of non-protein-coding genes and the malignancy of breast tumors are still unusual in the literature, making them interesting targets to study as well.

In another line of research of the group, focused on non-protein-coding genes, about 10 long non-protein-coding RNAs (IncRNAs) differentially expressed in human breast cancer cell lines were identified. One of them, LINC01133, was selected for functional characterization experiments, and an increase in malignancy was observed in the generated strains (Figure 1), both in the non-tumorigenic line MCF10A, in which we over-expressed this marker in an induced manner, and in the tumorigenic line Hs578T, in which we knocked out the marker via CRISPR/Cas9 gene-editing system (data not yet published). Therefore, the projects the group develops have the following objectives:

1) Identify and analyze the gene expression, via qRT-PCR, of new protein-coding and non-coding genes in a panel of human breast cancer cell lines, which present different degrees of malignancy;

2) Select new targets and functionally characterize them through gain and loss of function experiments to discover the relationship of these targets with the maintenance of normal cells and the progression of the tumor phenotype;

3) Functionally characterize the action mechanism of new targets in the tumor progression model by RNA-Seq and Western Blotting;

Figure 1: Analysis of invasive potential in Hs578T cells knocked out from lncRNA LINC01133. Representative photos referring to each cell line at the end of the experiment, in phase contrast field and developed with DAPI (Advanced Diagnosis Imaging).

REFERENCES

[1] BRAY, Freddie et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, v. 68, n. 6, p. 394-424, 2018.

[2] LOBBA, A. R. M. et al. Differential expression of CD90 and CD14 stem cell markers in malignant breast cancer cell lines. Cytometry Part A, v. 81, n. 12, p. 1084-1091, 2012.

[3] LOBBA, Aline Ramos Maia et al. High CD90 (THY-1) expression positively correlates with cell transformation and worse prognosis in basal-like breast cancer tumors. PloS one, v. 13, n. 6, p. e0199254, 2018.