Repositorio UFRO · Omeka S

Downstream process and evaluation of the concomitant impact of a recombinant glycosylated L-asparaginase on leukemic cancer cells and the bone marrow tumor microenvironment

Primer Autor	Calle, Yolanda
Co-autores	Kleingesinds, Eduardo Krebs Parizotto, Leticia de Almeida Effer, Brian Monteiro, Gisele Long, Paul F. Arroyo-Berdugo, Yoana Behrends, Volker Esposito, Maria Teresa Pessoa-Jr, Adalberto
Título	Downstream process and evaluation of the concomitant impact of a recombinant glycosylated L-asparaginase on leukemic cancer cells and the bone marrow tumor microenvironment
Editorial	ELSEVIER SCI LTD
Revista	PROCESS BIOCHEMISTRY
Lenguaje	en
Resumen	L-asparaginase (L-ASNase) is a life-saving medication used in the treatment of Acute Lymphoblastic Leukemia (ALL) which affects over 60,000 people yearly. However, L-ASNase still requires improvement since up to 60% of the patients develop hypersensitivity due to its immunogenicity. To address this issue, this work describes the downstream process of an L-ASNase from Erwinia chrysanthemi expressed extracellularly by Pichia pastoris with human-like glycosylation pattern and its further impact on leukemic cells co-cultured with bone marrow (BM) cytoprotective stromal cells. After size exclusion chromathography, a final yield of 54.93% was achieved and the proteomics analyses confirmed the attainment of an extremely pure enzyme. Glycosylated L-ASNase induced the complete inhibition of the proliferation of ALL and Acute Myeloid Leukemia (AML) cell lines tested, independently of the presence of BM stromal cells. However, the cytotoxic efficacy (induction of apoptosis) of glycosylated L-ASNase varied across cell lines, with ALL cell lines showing the most sensitivity. Additionally, the proapoptotic effect of glycosylated L-ASNase was partially inhibited by BM stromal cells. Taken together, our data warrant further investigations for the use of glycosylated L-ASNase against ALL and AML that should take into consideration the mechanisms of resistance mediated by the BM stroma for improved efficacy.
Fecha Publicación	2023
Tipo de Recurso	artículo original
doi	10.1016/j.procbio.2023.06.006
Formato Recurso	PDF
Palabras Claves	L-ASNase Leukemia Pichia pastoris Protein purification Cross-flow filtration Tumour microenvironment
Ubicación del archivo	http://dx.doi.org/10.1016/j.procbio.2023.06.006
Categoría OCDE	Bioquímica y Biología Molecular Biotecnología y Microbiología Aplicada Ingeniería
Materias	L-ASNasa Leucemia Pichia pastoris Purificación de proteínas Filtración de flujo cruzado Microambiente tumoral
Página de inicio (Recomendado-único)	41.0
Página final (Recomendado-único)	51
Identificador del recurso (Mandatado-único)	artículo original
Versión del recurso (Recomendado-único)	versión publicada
License	CC BY 4.0
Condición de la licencia (Recomendado-repetible)	CC BY 4.0
Derechos de acceso	acceso abierto
Access Rights	acceso abierto
Id de Web of Science	WOS:001028014900001
ISSN	1359-5113
Tipo de ruta	verde# hibrida
Categoría WOS	Bioquímica y Biología Molecular Biotecnología y Microbiología Aplicada Ingeniería
Referencia del Financiador (Mandatado si es aplicable-repetible)	ANID-FONDECYT 3210142 CONICYT 21150288 FAPESP 2013/08617-7 FAPESP 2015/07749-2 FAPESP 2017/20384-9 FAPESP 2017/25065-9 FAPESP 2018/15104-0 FAPESP 2018/03734-9 FAPESP 2019/06919-2 UFRO DI12-PEO1 EXE12-0004 CNPQ 309224/2019-5 CRUK C34579/A20784 ANID FONDECYT 3210142 ANID CONICYT 21150288 CNPq 309224/2019-5