Discovery of Novel Fetal Hemoglobin Inducers through Small Chemical Library Screening

Abstract

The screening of chemical libraries based on cellular biosensors is a useful approach to identify new hits for novel therapeutic targets involved in rare genetic pathologies, such as β-thalassemia and sickle cell disease. In particular, pharmacologically mediated stimulation of human γ-globin gene expression, and increase of fetal hemoglobin (HbF) production, have been suggested as potential therapeutic strategies for these hemoglobinopathies. In this article, we screened a small chemical library, constituted of 150 compounds, using the cellular biosensor K562.GR, carrying enhanced green fluorescence protein (EGFP) and red fluorescence protein (RFP) genes under the control of the human γ-globin and β-globin gene promoters, respectively. Then the identified compounds were analyzed as HbF inducers on primary cell cultures, obtained from β-thalassemia patients, confirming their activity as HbF inducers, and suggesting these molecules as lead compounds for further chemical and biological investigations.

Oncologia e Hematologia - Edição VI

“…Ao testar esses compostos em culturas primárias de células precursoras eritróides de pa ientes om β-talassemia, observou-se um aumento significativo na produção de hemoglobina fetal (HbF) em resposta ao tratamento. Esses resultados sugerem o potencial dessas moléculas como indutores químicos da síntese de HbF, oferecendo perspectivas promissoras para o desenvolvimento de tratamentos mais efi azes para ondições omo β-talassemia (BREVEGLIERI et al, 2020). Em outra pesquisa realizada relatou sobre dois pacientes portadores de anemia falciforme que foram tratados com com edição CRISPR-Cas9 do intensificador eritróide BCL11A para diminuir a expressão de BCL11A em células eritróides, após o tratamento, ambos os pacientes tiveram aumento de HbF de 43% por HPLC em um e 93% o outro.…”

Identification of small molecule agonists of fetal hemoglobin expression for the treatment of sickle cell disease

“…The systematic profiling of chemical agents to identify those capable of reactivating γ-globin expression has been hampered by the lack of suitable cell lines and methods. Recent reports of small molecule screens for fetal hemoglobin inducers have mainly employed reporter cell lines engineered with a partial γ-globin promoter driving a reporter gene, such as green fluorescent protein (GFP) or luciferase 21–24 . Although tractable for high-throughput compound screening, these synthetic gene constructs may not address essential endogenous DNA elements that tightly regulate and control γ-globin gene expression.…”