RESUMEN
Acute lymphoblastic leukemias (ALL) positive for KMT2A/AFF1 (MLL/AF4) translocation, which constitute 60% of all infant ALL cases, have a poor prognosis even after allogeneic hematopoietic stem cell transplantation (allo-HSCT). This poor prognosis is due to one of two factors, either resistance to TNFα, which mediates a graft-versus-leukemia (GVL) response after allo-HSCT, or immune resistance due to upregulated expression of the immune escape factor S100A6. Here, we report an immune stimulatory effect against KMT2A/AFF1-positive ALL cells by treatment with the anti-allergy drug amlexanox, which we found to inhibit S100A6 expression in the presence of TNF-α. In KMT2A/AFF1-positive transgenic (Tg) mice, amlexanox enhanced tumor immunity and lowered the penetrance of leukemia development. Similarly, in a NOD/SCID mouse model of human KMT2A/AFF1-positive ALL, amlexanox broadened GVL responses and extended survival. Our findings show how amlexanox degrades the resistance of KMT2A/AFF1-positive ALL to TNFα by downregulating S100A6 expression, with immediate potential implications for improving clinical management of KMT2A/AFF1-positive ALL. Cancer Res; 77(16); 4426-33. ©2017 AACR.
Asunto(s)
Aminopiridinas/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Proteínas de Ciclo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Proteínas S100/metabolismo , Factores de Elongación Transcripcional/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Animales , Antialérgicos/administración & dosificación , Antialérgicos/farmacología , Proteínas de Ciclo Celular/genética , Proteínas de Unión al ADN/genética , Modelos Animales de Enfermedad , Regulación hacia Abajo , Sinergismo Farmacológico , Humanos , Masculino , Ratones , Ratones Endogámicos NOD , Ratones SCID , Ratones Transgénicos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Proteína A6 de Unión a Calcio de la Familia S100 , Proteínas S100/genética , Factores de Elongación Transcripcional/genética , Factor de Necrosis Tumoral alfa/administración & dosificaciónRESUMEN
Mixed-lineage leukemia (MLL)/AF4-positive acute lymphoblastic leukemia (ALL) is a common type of leukemia in infants, which is associated with a high relapse rate and poor prognosis. IL24 selectively induces apoptosis in cancer cells and exerts immunomodulatory and antiangiogenic effects. We examined the effects of adeno-associated virus type 8 (AAV8) vector-mediated muscle-directed systemic gene therapy in MLL/AF4-positive ALL using IL24. In a series of in vitro studies, we examined the effects of AAV8-IL24-transduced C2C12 cell-conditioned medium. We also examined the effects of AAV8-IL24 in MLL/AF4 transgenic mice. The results revealed the effects of AAV8-IL24 in MLL/AF4-positive ALL both in vitro and in vivo. With regard to the mechanism of therapy using AAV8-IL24 in MLL/AF4-positive ALL, we demonstrated the antiangiogenicity and effects on the ER stress pathway and unreported pathways through inhibition of S100A6 and HOXA9, which is specific to MLL/AF4-positive ALL. Inhibition of S100A6 by IL24 was dependent on TNF-α and induced acetylation of p53 followed by activation of the caspase 8-caspase 3 apoptotic pathway. Inhibition of HOXA9 by IL24, which was independent of TNF-α, induced MEIS1 activation followed by activation of the caspase 8-caspase 3 apoptotic pathway. Thus, gene therapy using AAV8-IL24 is a promising treatment for MLL/AF4-positive ALL.
Asunto(s)
Dependovirus/genética , Modelos Animales de Enfermedad , Terapia Genética , Interleucinas/genética , Proteína de la Leucemia Mieloide-Linfoide/antagonistas & inhibidores , Proteína de la Leucemia Mieloide-Linfoide/genética , Proteínas de Fusión Oncogénica/antagonistas & inhibidores , Proteínas de Fusión Oncogénica/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Animales , Apoptosis , Western Blotting , Proteínas de Ciclo Celular/metabolismo , Proliferación Celular , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/citología , Fibroblastos/metabolismo , Citometría de Flujo , Proteínas de Homeodominio/metabolismo , Humanos , Técnicas para Inmunoenzimas , Masculino , Ratones , Ratones Transgénicos , Mioblastos/citología , Mioblastos/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , ARN Mensajero/genética , Proteína A6 de Unión a Calcio de la Familia S100 , Proteínas S100/metabolismoRESUMEN
The purpose of this study was to investigate which of the three readily available scaffold materials would be suitable for adipose tissue engineering when implanted with adipose-derived stem cells (ASCs) in vivo. ASCs isolated from green fluorescence protein (GFP) transgenic mice were incubated in an adipogenic medium and then seeded onto type I collagen sponge, non-woven polyglycolic acid or hyaluronic acid gel. The constructs were harvested and evaluated histologically and immunohistochemically 4 and 8 weeks after subcutaneous implantation into athymic mice. The gene expression of peroxisome-proliferator-activated receptor gamma2 (PPAR-gamma2), the adipocyte-specific transcriptional factor, was also investigated by using reverse transcription-polymerase chain reaction. Histological examination showed that more adipose-tissue-like construct was regenerated when using type I collagen sponge than when the other scaffolds were used. Moreover, immunohistostaining revealed that some of the adipocytes on the type I collagen construct expressed GFP. PPAR-gamma2 gene expression in the induced ASCs in the type I collagen sponge was observed. These findings suggest that type I collagen sponge may be the most suitable among the three readily available scaffolds for adipogenesis.
Asunto(s)
Tejido Adiposo/citología , Trasplante de Células Madre/métodos , Células Madre/citología , Ingeniería de Tejidos/métodos , Andamios del Tejido , Adipogénesis/genética , Tejido Adiposo/metabolismo , Animales , Diferenciación Celular , Femenino , Expresión Génica , Inmunohistoquímica , Ratones , Ratones Transgénicos , PPAR gamma/biosíntesis , PPAR gamma/genética , ARN/biosíntesis , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Madre/metabolismoRESUMEN
P230 Bcr/Abl has been associated with indolent myeloproliferative disease (MPD). We generated transgenic mice expressing P230Bcr/Abl driven by the promoter of the long terminal repeat of the murine stem cell virus of the MSCV neo P230 BCR/ABL vector. Two founder mice exhibited mild granulocytosis and marked thrombocytosis and developed MPD. The disease of one founder mouse, no. 13, progressed to extramedullary myeloblastic crisis in the liver at 12 months old. The other founder mouse, no. 22, was found to have chronic-phase MPD with large populations of megakaryocytes and granulocytes in an enlarged spleen. The transgenic progeny of no. 22 clearly exhibited MPD at 15 months old. These results showed that P230Bcr/Abl had leukemogenic properties and induced MPD. The phenotype of the MPD caused by P230Bcr/Abl was characterized by mild granulocytosis, a high platelet count, infiltration of megakaryocytes in some organs, and a longer disease latency compared with the MPD caused by P210Bcr/Abl.