Use of HSC Targeted LNP to Generate a Mouse Model of Lethal α-Thalassemia and Treatment via Lentiviral Gene Therapy.
Maxwell Elliott ChappellLaura BredaLucas TricoliAmaliris GuerraDanuta Jadwiga JarochaCarlo Castruccio CastracaniTyler Ellis PappNaoto TanakaNolan HamiltonMichael Paul TriebwasserValentina GhiaccioMegan FedorkyKandace GollompVeronica BochenekAoife M RocheJohn K EverettEmma J CookFrederic D BushmanNattiya TeawtrakulStavros GlentisAntonios KattamisBarbara L MuiYing K TamDrew WeissmanOsheiza AbdulmalikHamideh ParhizStefano RivellaPublished in: Blood (2024)
-Thalassemia (AT) is one of the most commonly occurring inherited hematological diseases. However, few treatments are available, and allogeneic bone marrow transplantation (BMT) is the only available therapeutic option for patients with severe AT. Research into AT has remained limited due to a lack of adult mouse models, with severe AT typically resulting in in utero lethality. By using a lipid nanoparticle (LNP) targeting the receptor CD117 and delivering a Cre mRNA (mRNACreLNPCD117), we were able to delete floxed -globin genes at high efficiency in hematopoietic stem cells (HSC) ex vivo. These cells were then engrafted in the absence or presence of a novel α-globin expressing lentiviral vector (ALS20I). Myeloablated mice transplanted with mRNACreLNPCD117-treated HSC showed a complete knockout of -globin genes. They demonstrated a phenotype characterized by the synthesis of hemoglobin H (-tetramers, or HbH), aberrant erythropoiesis, and abnormal organ morphology, culminating in lethality approximately eight weeks following engraftment. Mice receiving mRNACreLNPCD117-treated HSC with at least one copy of ALS20I survived long-term with normalization of erythropoiesis, decreased the production of HbH, and ameliorated the abnormal organ morphology. Furthermore, we tested ALS20I in erythroid progenitors derived from -globin-KO CD34+ and cells isolated from patients with both deletional and non-deletional HbH disease, demonstrating improvement in -globin/-globin mRNA ratio and reduction in the formation of HbH by HPLC. Our results demonstrate the broad applicability of LNP for disease modeling, characterization of a novel severe mouse model of AT, and the efficacy of ALS20I for treating AT.
Keyphrases
- mouse model
- gene therapy
- bone marrow
- induced apoptosis
- stem cells
- amyotrophic lateral sclerosis
- high efficiency
- cell cycle arrest
- early onset
- mesenchymal stem cells
- genome wide
- high fat diet induced
- cancer therapy
- wild type
- binding protein
- cell therapy
- gene expression
- type diabetes
- newly diagnosed
- hematopoietic stem cell
- sickle cell disease
- cell death
- skeletal muscle
- drug delivery
- gestational age
- insulin resistance
- high dose
- genome wide analysis
- replacement therapy