Background:𝛽-hemoglobinopathies are one of the most common recessive genetic diseases worldwide,with limited treatments available,particularly in developed countries where the prevalence is higher.Pharmacologi...Background:𝛽-hemoglobinopathies are one of the most common recessive genetic diseases worldwide,with limited treatments available,particularly in developed countries where the prevalence is higher.Pharmacological reactivation of Fetal Hemoglobin(HbF)is a promising therapeutic strategy.However,approximately 25%of the patients do not respond to Hydroxyurea(HU),the first and most commonly used HbF inducing agent approved by the FDA.Objective:Here,we performed an in vitro assessment of transcriptional effects induced by natural bioactive compounds,namely Epigallocatechin-3-gallate(EGCG)and genistein(GN)in globin genes(HBA1,HBB,HBG1 and HBG2)in HbF regulators/silencer genes(KLF1,BCL11A,MYB and BGLT3)and in epigenetic regulator genes(DNMT1,DNMT3A,DNMT3B,HDAC1,HDAC2,HDAC3 and HDAC8).Moreover,we evaluated EGCG’s in vivo effects in hematological parameters of healthy volunteers.Methods:K562 cells were exposed for 72 and 96 h to GN and EGCG at 100,250 and 500 ng/mL.Cell proliferation and viability were measured,and transcriptional levels were evaluated by qRT-PCR.For in vivo assay,complete blood count was determined by flow cytometry and HbF level was determined through HPLC in 30 healthy individuals before and after 225 mg EGCG ingestion per day during a 90-day period.Results:Both compounds impact cellular metabolism and proliferation with no cytotoxic effects.Divergent GN and EGCG effects in globin and BGLT3 expression levels suggest the involvement of divergent signaling pathways.As for the epigenetic potential,EGCG particularly affects HDAC2 and HDAC8 transcription,whereas GN signifi-cantly affects expression patterns of methylation and acetylation modulators.HU appears to have time divergent effects,with greater impact in methylation at 72 h(overregulates DNTM3A)while affecting acetylation mostly at 96 h(downregulates HDAC1 and HDAC8).Additionally,in vivo,EGCG demonstrated a modulator effect in hematopoiesis and HbF induction.Conclusion:Our results advocate EGCG and GN with HbF pharmacological reactivation potential and sustain further research as new alternative approaches for𝛽-hemoglobinopathies therapies.展开更多
With advancements in gene editing technologies,our ability to make precise and efficient modifications to the genome is increasing at a remarkable rate,paving the way for scientists and clinicians to uniquely treat a ...With advancements in gene editing technologies,our ability to make precise and efficient modifications to the genome is increasing at a remarkable rate,paving the way for scientists and clinicians to uniquely treat a multitude of previously irremediable diseases.CRISPR-Cas9,short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9,is a gene editing platform with the ability to alter the nucleotide sequence of the genome in living cells.This technology is increasing the number and pace at which new gene editing treatments for genetic disorders are moving toward the clinic.Theβ-hemoglobinopathies are a group of monogenic diseases,which despite their high prevalence and chronic debilitating nature,continue to have few therapeutic options available.In this review,we will discuss our existing comprehension of the genetics and current state of treatment forβ-hemoglobinopathies,consider potential genome editing therapeutic strategies,and provide an overview of the current state of clinical trials using CRISPR-Cas9 gene editing.展开更多
目的探讨出生时胎龄<32周早产儿生后早期动脉血胎儿血红蛋白(HbF)水平与早产儿视网膜病(retinopathy of prematurity,ROP)发病的关系。方法选择2019年3月—2022年2月我院新生儿科收治的出生时胎龄<32周的早产儿99例,根据是否诊断...目的探讨出生时胎龄<32周早产儿生后早期动脉血胎儿血红蛋白(HbF)水平与早产儿视网膜病(retinopathy of prematurity,ROP)发病的关系。方法选择2019年3月—2022年2月我院新生儿科收治的出生时胎龄<32周的早产儿99例,根据是否诊断为ROP,分为ROP组(34例)和非ROP组(65例)。比较两组患儿的临床资料,采用Logistic回归法分析患儿生后2周动脉血HbF水平与ROP的相关性。结果ROP组患儿出生时胎龄以及出生体质量均显著低于非ROP组(t=3.371、3.218,P<0.05),出生时胎龄<28周患儿构成比、出生体质量<1000 g患儿构成比、顺产患儿构成比、生后2周内输血患儿构成比,以及用氧时间与住院时间均显著高于或长于非ROP组(χ^(2)=4.390~11.579,Z=3.283、5.207,P<0.05)。ROP组患儿生后2周动脉血血红蛋白(Hb)水平及HbF比例均显著低于非ROP组(t=-2.791、-3.599,P<0.05)。Logistic回归法分析显示,动脉血HbF比例降低是ROP发生的危险因素(OR=0.915,95%CI=0.849~0.987,P<0.05)。结论出生时胎龄<32周早产儿生后2周动脉血HbF比例降低是ROP发生的危险因素,维持较高的动脉血HbF水平可能对临床ROP的发生具有预防作用。展开更多
基金an IDI&CA grant IPL/2019/HemoFet_ESTeSL and by H&TRC-Health&Technology Re-search Center,ESTeSL-Escola Superior de Tecnologia da Saúde,Insti-tuto Politécnico de Lisboa.
文摘Background:𝛽-hemoglobinopathies are one of the most common recessive genetic diseases worldwide,with limited treatments available,particularly in developed countries where the prevalence is higher.Pharmacological reactivation of Fetal Hemoglobin(HbF)is a promising therapeutic strategy.However,approximately 25%of the patients do not respond to Hydroxyurea(HU),the first and most commonly used HbF inducing agent approved by the FDA.Objective:Here,we performed an in vitro assessment of transcriptional effects induced by natural bioactive compounds,namely Epigallocatechin-3-gallate(EGCG)and genistein(GN)in globin genes(HBA1,HBB,HBG1 and HBG2)in HbF regulators/silencer genes(KLF1,BCL11A,MYB and BGLT3)and in epigenetic regulator genes(DNMT1,DNMT3A,DNMT3B,HDAC1,HDAC2,HDAC3 and HDAC8).Moreover,we evaluated EGCG’s in vivo effects in hematological parameters of healthy volunteers.Methods:K562 cells were exposed for 72 and 96 h to GN and EGCG at 100,250 and 500 ng/mL.Cell proliferation and viability were measured,and transcriptional levels were evaluated by qRT-PCR.For in vivo assay,complete blood count was determined by flow cytometry and HbF level was determined through HPLC in 30 healthy individuals before and after 225 mg EGCG ingestion per day during a 90-day period.Results:Both compounds impact cellular metabolism and proliferation with no cytotoxic effects.Divergent GN and EGCG effects in globin and BGLT3 expression levels suggest the involvement of divergent signaling pathways.As for the epigenetic potential,EGCG particularly affects HDAC2 and HDAC8 transcription,whereas GN signifi-cantly affects expression patterns of methylation and acetylation modulators.HU appears to have time divergent effects,with greater impact in methylation at 72 h(overregulates DNTM3A)while affecting acetylation mostly at 96 h(downregulates HDAC1 and HDAC8).Additionally,in vivo,EGCG demonstrated a modulator effect in hematopoiesis and HbF induction.Conclusion:Our results advocate EGCG and GN with HbF pharmacological reactivation potential and sustain further research as new alternative approaches for𝛽-hemoglobinopathies therapies.
文摘With advancements in gene editing technologies,our ability to make precise and efficient modifications to the genome is increasing at a remarkable rate,paving the way for scientists and clinicians to uniquely treat a multitude of previously irremediable diseases.CRISPR-Cas9,short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9,is a gene editing platform with the ability to alter the nucleotide sequence of the genome in living cells.This technology is increasing the number and pace at which new gene editing treatments for genetic disorders are moving toward the clinic.Theβ-hemoglobinopathies are a group of monogenic diseases,which despite their high prevalence and chronic debilitating nature,continue to have few therapeutic options available.In this review,we will discuss our existing comprehension of the genetics and current state of treatment forβ-hemoglobinopathies,consider potential genome editing therapeutic strategies,and provide an overview of the current state of clinical trials using CRISPR-Cas9 gene editing.
文摘目的探讨出生时胎龄<32周早产儿生后早期动脉血胎儿血红蛋白(HbF)水平与早产儿视网膜病(retinopathy of prematurity,ROP)发病的关系。方法选择2019年3月—2022年2月我院新生儿科收治的出生时胎龄<32周的早产儿99例,根据是否诊断为ROP,分为ROP组(34例)和非ROP组(65例)。比较两组患儿的临床资料,采用Logistic回归法分析患儿生后2周动脉血HbF水平与ROP的相关性。结果ROP组患儿出生时胎龄以及出生体质量均显著低于非ROP组(t=3.371、3.218,P<0.05),出生时胎龄<28周患儿构成比、出生体质量<1000 g患儿构成比、顺产患儿构成比、生后2周内输血患儿构成比,以及用氧时间与住院时间均显著高于或长于非ROP组(χ^(2)=4.390~11.579,Z=3.283、5.207,P<0.05)。ROP组患儿生后2周动脉血血红蛋白(Hb)水平及HbF比例均显著低于非ROP组(t=-2.791、-3.599,P<0.05)。Logistic回归法分析显示,动脉血HbF比例降低是ROP发生的危险因素(OR=0.915,95%CI=0.849~0.987,P<0.05)。结论出生时胎龄<32周早产儿生后2周动脉血HbF比例降低是ROP发生的危险因素,维持较高的动脉血HbF水平可能对临床ROP的发生具有预防作用。
