Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration
Authors
FEDOROVÁ, Veronika (203 Czech Republic, belonging to the institution), Veronika POSPÍŠILOVÁ (203 Czech Republic, belonging to the institution), Tereza VÁŇOVÁ (203 Czech Republic, belonging to the institution), Kateřina AMRUZ ČERNÁ (203 Czech Republic, belonging to the institution), Pavel ABAFFY (703 Slovakia), Jiří SEDMÍK (203 Czech Republic, belonging to the institution), Jan RAŠKA (203 Czech Republic, belonging to the institution), Simona VOCHYÁNOVÁ (203 Czech Republic, belonging to the institution), Zuzana MATŮŠOVÁ (203 Czech Republic), Jana HOUSEROVÁ (203 Czech Republic), Lukáš VALIHRACH (203 Czech Republic), Zdeněk HODNÝ (203 Czech Republic) and Dáša BOHAČIAKOVÁ (703 Slovakia, guarantor, belonging to the institution)
It is currently challenging to adequately model the growth and migration of glioblastoma using two-dimensional (2D) in vitro culture systems as they quickly lose the original, patient-specific identity and heterogeneity. However, with the advent of three-dimensional (3D) cell cultures and human-induced pluripotent stem cell (iPSC)-derived cerebral organoids (COs), studies demonstrate that the glioblastoma-CO (GLICO) coculture model helps to preserve the phenotype of the patient-specific tissue. Here, we aimed to set up such a model using mature COs and develop a pipeline for subsequent analysis of cocultured glioblastoma. Our data demonstrate that the growth and migration of the glioblastoma cell line within the mature COs are significantly increased in the presence of extracellular matrix proteins, shortening the time needed for glioblastoma to initiate migration. We also describe in detail the method for the visualization and quantification of these migrating cells within the GLICO model. Lastly, we show that this coculture model (and the human brain-like microenvironment) can significantly transform the gene expression profile of the established U87 glioblastoma cell line into proneural and classical glioblastoma cell types.
Links
EF16_013/0001761, research and development project
Name: RECETOX RI
EF17_043/0009632, research and development project
Name: CETOCOEN Excellence
EF19_073/0016943, research and development project
Name: Interní grantová agentura Masarykovy univerzity
GA20-15728S, research and development project
Name: Nové přístupy k překonání terapeutické rezistence u glioblastoma multiforme se zaměřením na buněčnou senescenci a model mozkového organoidu
Investor: Czech Science Foundation
GA21-21510S, research and development project
Name: Studium molekulárních mechanismů vzniku Alzheimerovy choroby pomocí cerebrálních organoidů (Acronym: AD Brain Organoids)
Investor: Czech Science Foundation
GJ18-25429Y, research and development project
Name: Funkční studie mikroRNA u nerálních kmenových buněk v průběhu diferenciace
Investor: Ministry of Education, Youth and Sports of the CR, RECETOX RI
LM2018129, research and development project
Name: Národní infrastruktura pro biologické a medicínské zobrazování Czech-BioImaging
Investor: Ministry of Education, Youth and Sports of the CR
LM2018132, research and development project
Name: Národní centrum lékařské genomiky (Acronym: NCLG)
Investor: Ministry of Education, Youth and Sports of the CR, National Center for Medical Genomics
MUNI/A/1398/2021, interní kód MU
Name: Zdroje pro tkáňové inženýrství 12 (Acronym: TissueEng 12)
Investor: Masaryk University
MUNI/IGA/1273/2021, interní kód MU
Name: Investigation of the role of candidate miRNAs in neural stem cells’ identity
Investor: Masaryk University
MUNI/R/1321/2021, interní kód MU
Name: Career Restart - Next generation sequencing in neural development and disease
Investor: Masaryk University, CAREER RESTART
MUNI/R/1697/2020, interní kód MU
Name: Career Restart - Stem Cell-Based Models of Neurodegeneration
Investor: Masaryk University, CAREER RESTART
MUNI/11/SUP/01/2022, interní kód MU
Name: Role miRNA v šíření Glioblastoma multiforme mozkovou tkání in vitro
Investor: Masaryk University, Start-up
MUNI/11/SUP/19/2020, interní kód MU
Name: Efekt buněčného stárnutí na vznik Alzheimerovy nemoci
Investor: Masaryk University
NU21-08-00373, research and development project
Name: Patogeny indukovaná senescence jako spouštěcí faktor Alzheimerovy nemoci (Acronym: Patogeny u AD)
Investor: Ministry of Health of the CR, Subprogram 1 - standard
NV19-08-00472, research and development project
Name: Klinicky relevantní biochemické, imunologické a buněčné biomarkery Alzheimerovy nemoci a stárnutí (Acronym: BioMAD)
Investor: Ministry of Health of the CR
ROZV/23/LF19/2019, interní kód MU
Name: Využití in vitro kultivovaných cerebrálních organoidů pro modelování patogeneze Alzheimerovy choroby
Investor: Ministry of Education, Youth and Sports of the CR, Internal development projects
ROZV/28/LF22/2020, interní kód MU
Name: ln vitro modelováni Alzheimerovy choroby pomoci nově derivovaných indukovaných plunpotentnich kmenových buněk z pacientu se spontánní formou onemocnění.
Investor: Ministry of Education, Youth and Sports of the CR, Internal development projects
FEDOROVÁ, Veronika, Veronika POSPÍŠILOVÁ, Tereza VÁŇOVÁ, Kateřina AMRUZ ČERNÁ, Pavel ABAFFY, Jiří SEDMÍK, Jan RAŠKA, Simona VOCHYÁNOVÁ, Zuzana MATŮŠOVÁ, Jana HOUSEROVÁ, Lukáš VALIHRACH, Zdeněk HODNÝ and Dáša BOHAČIAKOVÁ. Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration. MOLECULAR ONCOLOGY. ENGLAND: WILEY, 2023, vol. 17, No 4, p. 647-663. ISSN 1574-7891. Available from: https://dx.doi.org/10.1002/1878-0261.13389.
@article{2263677, author = {Fedorová, Veronika and Pospíšilová, Veronika and Váňová, Tereza and Amruz Černá, Kateřina and Abaffy, Pavel and Sedmík, Jiří and Raška, Jan and Vochyánová, Simona and Matůšová, Zuzana and Houserová, Jana and Valihrach, Lukáš and Hodný, Zdeněk and Bohačiaková, Dáša}, article_location = {ENGLAND}, article_number = {4}, doi = {http://dx.doi.org/10.1002/1878-0261.13389}, keywords = {cerebral organoids; GLICO; glioblastoma; induced pluripotent stem cells}, language = {eng}, issn = {1574-7891}, journal = {MOLECULAR ONCOLOGY}, title = {Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration}, url = {https://febs.onlinelibrary.wiley.com/doi/10.1002/1878-0261.13389}, volume = {17}, year = {2023} }
TY - JOUR ID - 2263677 AU - Fedorová, Veronika - Pospíšilová, Veronika - Váňová, Tereza - Amruz Černá, Kateřina - Abaffy, Pavel - Sedmík, Jiří - Raška, Jan - Vochyánová, Simona - Matůšová, Zuzana - Houserová, Jana - Valihrach, Lukáš - Hodný, Zdeněk - Bohačiaková, Dáša PY - 2023 TI - Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration JF - MOLECULAR ONCOLOGY VL - 17 IS - 4 SP - 647-663 EP - 647-663 PB - WILEY SN - 15747891 KW - cerebral organoids KW - GLICO KW - glioblastoma KW - induced pluripotent stem cells UR - https://febs.onlinelibrary.wiley.com/doi/10.1002/1878-0261.13389 N2 - It is currently challenging to adequately model the growth and migration of glioblastoma using two-dimensional (2D) in vitro culture systems as they quickly lose the original, patient-specific identity and heterogeneity. However, with the advent of three-dimensional (3D) cell cultures and human-induced pluripotent stem cell (iPSC)-derived cerebral organoids (COs), studies demonstrate that the glioblastoma-CO (GLICO) coculture model helps to preserve the phenotype of the patient-specific tissue. Here, we aimed to set up such a model using mature COs and develop a pipeline for subsequent analysis of cocultured glioblastoma. Our data demonstrate that the growth and migration of the glioblastoma cell line within the mature COs are significantly increased in the presence of extracellular matrix proteins, shortening the time needed for glioblastoma to initiate migration. We also describe in detail the method for the visualization and quantification of these migrating cells within the GLICO model. Lastly, we show that this coculture model (and the human brain-like microenvironment) can significantly transform the gene expression profile of the established U87 glioblastoma cell line into proneural and classical glioblastoma cell types. ER -
FEDOROVÁ, Veronika, Veronika POSPÍŠILOVÁ, Tereza VÁŇOVÁ, Kateřina AMRUZ ČERNÁ, Pavel ABAFFY, Jiří SEDMÍK, Jan RAŠKA, Simona VOCHYÁNOVÁ, Zuzana MATŮŠOVÁ, Jana HOUSEROVÁ, Lukáš VALIHRACH, Zdeněk HODNÝ and Dáša BOHAČIAKOVÁ. Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration. \textit{MOLECULAR ONCOLOGY}. ENGLAND: WILEY, 2023, vol.~17, No~4, p.~647-663. ISSN~1574-7891. Available from: https://dx.doi.org/10.1002/1878-0261.13389.