PML/RARa leukemia induced murine model for immunotherapy evaluation

Autor(es)

Shiraishi, Rodrigo N. ; Bombeiro, Andre L. ; Castro, Tamara C. L. ; Della Via, Fernanda I. ; Santos, Irene ; Rego, Eduardo M. ; Saad, Sara T. O. ; Torello, Cristiane O.

• Tipo de documento: Artigo Científico
• Fonte: TRANSPLANT IMMUNOLOGY; v. 81, p. 14-pg., 2023-08-26.

Resumo

Even though leukemia murine models are valuable tools for new drug therapy studies, most of these models consist of immunocompromised mice, which do not exhibit immune responses. In order to obtain an adequate leukemia model, we established an acute promyelocytic leukemia transplantation-based model (PML/RARa) in immunocompetent BALB/c mice, thus making it possible to study drug-induced cellular immune responses in leukemia. The development of PML/RARa leukemia was confirmed by leukocytosis (76.27 +/- 21.8 vs. 3.40 +/- 1.06; P < 0.0001), anemia (7.46 +/- 1.86 vs. 15.10 +/- 0.96; P < 0.0001), and thrombocytopenia (131.85 +/- 39.32 vs. 839.50 +/- 171.20; P < 0.0001), and the presence of blasts in the peripheral blood of mice (approximately 50% blasts; P < 0.0001), 15 days after the transplants. These findings were corroborated through differential counts, flow cytometry, and in vivo imaging, which indicated increased number of immature cells in the bone marrow (15.75 +/- 3.30 vs 6.69 +/- 0.55; P < 0.001), peripheral blood (7.88 +/- 2.67 vs 1.22 +/- 0.89; P < 0.001), and spleen (35.21 +/- 4.12 vs 1.35 +/- 0.86; P < 0.0001), as well as promyelocytes in the bone marrow (41.23 +/- 4.80 vs 5.73 +/- 1.50; P < 0.0001), peripheral blood (46.08 +/- 7.52 vs 1.10 +/- 0.59; P < 0.0001) and spleen (35.31 +/- 8.26 vs 2.49 +/- 0.29; P < 0.0001) of PML/RARa mice. Compared to basal conditions of untransplanted mice, the PML/ RARa mice exhibited frequencies of T lymphocytes CD4 helper = 14.85 +/- 2.91 vs 20.77 +/- 2.9 in the peripheral blood (P < 0.05); 12.75 +/- 1.33 vs 45.90 +/- 2.02 in the spleen (P < 0.0001); CD8 cytotoxic = 11.27 +/- 3.44 vs 11.05 +/- 1.22 in the peripheral blood (P > 0.05); 10.48 +/- 1.16 vs 30.02 +/- 1.80 in the spleen (P < 0.0001); natural killer (NK) cells = 3.68 +/- 1.35 vs 6.84 +/- 0.52 in the peripheral blood (P < 0.001); 4.43 +/- 0.57 vs 6.40 +/- 1.14 in the spleen (P < 0.05); B cells 2.50 +/- 0.60 vs 15.20 +/- 5.34 in the peripheral blood (P < 0.001); 17.77 +/- 4.39 vs 46.90 +/- 5.92 in the spleen (P < 0.0001); neutrophils = 5.97% +/- 1.88 vs 31.57 +/- 9.14 (P < 0.0001); and monocytes = 6.45 +/- 2.97 vs 15.85 +/- 2.57 (P < 0.001), selected as classical (3.33 +/- 3.40 vs 57.80 +/- 16.51, P < 0.0001), intermediate (57.42 +/- 10.61 vs 21.75 +/- 5.90, P < 0.0001), and non-classical monocytes (37.51 +/- 10.85 vs 18.08 +/- 7.13, P < 0.05) in the peripheral blood; and as classically activated (M1) within in the bone marrow (3.70 +/- 0.94 vs 1.88 +/- 0.39, P < 0.05) and spleen 15.19 +/- 3.32 vs 9.47 +/- 1.61, P < 0.05), in addition to alternatively activated (M2) macrophages within the bone marrow (23.06 +/- 5.25 vs 1.76 +/- 0.74, P < 0.0001) and spleen (46.51 +/- 11.18 vs 30.58 +/- 2.64, P < 0.05) compartments. YAll-trans retinoic acid (ATRA) treatment of PML/RARa mice reduced blast (immature cells) in the bone marrow (8.62 +/- 1.81 vs 15.76 +/- 1.25; P < 0.05) and spleen (8.75 +/- 1.31 vs 35.21 +/- 1.55; P < 0.0001) with no changes in the peripheral blood (10.13 +/- 3.33 vs 7.88 +/- 1.01; P > 0.05), as well as reduced promyelocytes in the bone marrow (19.79 +/- 4.84 vs 41.23 +/- 1.81; P < 0.05), peripheral blood (31.65 +/- 3.92 vs 46.09 +/- 2.84; P < 0.05) and spleen (24.84 +/- 2.03 vs 41.46 +/- 2.39; P < 0.001), and increased neutrophils of the peripheral blood (35.48 +/- 7.24 vs 7.83 +/- 1.40; P < 0.05) which was corroborated by reducing of immature cells and increase of neutrophil in the stained smears from PML/RARa mice, thus confirming that this model can be used in drug development studies. Our results show the effective induction of PML/RARa leukemia in BALB/c mice, thus producing a low-priced and reliable tool for investigating cellular immune responses in leukemia. (AU)

• Processo FAPESP: 13/21539-5 - Investigação dos efeitos do extrato de Rhodiola rosea e de chá verde (Camellia sinensis) em animais transplantados com a linhagem leucêmica U937
• Beneficiário: Rodrigo Naoto Shiraishi
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 17/21801-2 - Preditores de gravidade e novos tratamentos para neoplasias da medula óssea
• Beneficiário: Sara Teresinha Olalla Saad
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 11/51959-0 - Biologia das doenças neoplásicas da medula óssea
• Beneficiário: Sara Teresinha Olalla Saad
• Modalidade de apoio: Auxílio à Pesquisa - Temático