The role of Ese-2, an Ets family transcription factor, in gene regulation is not known. In this study, the interactionbetween Ese-2 and cytokeratin 18 (K18) intron 1 was characterized in lung epithelial cells. Reporte...The role of Ese-2, an Ets family transcription factor, in gene regulation is not known. In this study, the interactionbetween Ese-2 and cytokeratin 18 (K18) intron 1 was characterized in lung epithelial cells. Reporter gene assays showedEse-2 was able to upregulate K18 intron 1 enhanced reporter gene expression by approximately 2-fold. We found thatfull length Ese-2 did not bind DNA strongly, therefore truncated versions of the protein, containing the ETS domain orPointed domain, were created and tested in electrophoresis mobility shift assays. Multiple interactions between the ETSdomain and putative DNA binding sites within K18 intron 1 were observed, which led to the determination of a possibleEse-2 DNA binding consensus sequence. These experiments suggest that Ese-2 could play a role in the regulation ofK18 expression in lung epithelial cells.展开更多
Vision loss or impairment resulting from the degeneration of the retinal pigment epithelium and photoreceptor death affects millions worldwide.Recent exciting results from clinical studies of small numbers of patients...Vision loss or impairment resulting from the degeneration of the retinal pigment epithelium and photoreceptor death affects millions worldwide.Recent exciting results from clinical studies of small numbers of patients treated with human embryonic stem cell-derived retinal pigment epithelial cells may provide hope for affected individuals.展开更多
Asthma is a chronic lung disease that primarily affects the lower respiratory tract in more than 300 million people worldwide.Asthma is often triggered by exposure to certain substances or conditions,such as allergens...Asthma is a chronic lung disease that primarily affects the lower respiratory tract in more than 300 million people worldwide.Asthma is often triggered by exposure to certain substances or conditions,such as allergens or cold air.The overreaction to the allergens causes inflammation and narrowing of the airway,which leads to episodic or persistent symptoms including shortness of breath,nocturnal cough,chest tightness,and wheezing.展开更多
After two decades of ups and downs,gene therapy has recently achieved a milestone in treating patients with Leber’s congenital amaurosis(LCA).LCA is a group of inherited blinding diseases with retinal degeneration an...After two decades of ups and downs,gene therapy has recently achieved a milestone in treating patients with Leber’s congenital amaurosis(LCA).LCA is a group of inherited blinding diseases with retinal degeneration and severe vision loss in early infancy.Mutations in several genes,including RPE65,cause the disease.Using adenoassociated virus as a vector,three independent teams of investigators have recently shown that RPE65 can be delivered to retinal pigment epithelial cells of LCA patients by subretinal injections resulting in clinical benefits without side effects.However,considering the whole field of gene therapy,there are still major obstacles to clinical applications for other diseases.These obstacles include innate and immune barriers to vector delivery,toxicity of vectors and the lack of sustained therapeutic gene expression.Therefore,new strategies are needed to overcome these hurdles for achieving safe and effective gene therapy.In this article,we shall review the major advancements over the past two decades and,using lung gene therapy as an example,discuss the current obstacles and possible solutions to provide a roadmap for future gene therapy research.展开更多
Innate immune responses form the first line of defense against foreign insults and recently significant advances have been made in our understanding of the initiation of innate immune response along with its ability t...Innate immune responses form the first line of defense against foreign insults and recently significant advances have been made in our understanding of the initiation of innate immune response along with its ability to modulate inflammation. In airway diseases such as asthma, COPD and cystic fibrosis, over reacting of the airway innate immune responses leads to cytokine imbalance and airway remodeling or damage. Helper-dependent adenoviral vectors have the potential to deliver genes to modulate airway innate immune responses and have many advantages over its predecessors. However, there still are a few limitations that need to be addressed prior to their use in clinical applications.展开更多
Gene therapy has been considered as the most ideal medical intervention for genetic diseases because it is intended to target the cause of diseases instead of disease symptoms.Availability of techniques for identifica...Gene therapy has been considered as the most ideal medical intervention for genetic diseases because it is intended to target the cause of diseases instead of disease symptoms.Availability of techniques for identification of genetic mutations and for in vitro manipulation of genes makes it practical and attractive.After the initial hype in 1990s and later disappointments in clinical trials formore than a decade,light has finally come into the tunnel in recent years,especially in the field of eye gene therapy where it has taken big strides.Clinical trials in gene therapy for retinal degenerative diseases such as Leber’s congenital amaurosis(LCA)and choroideremia demonstrated clear therapeutic efficacies without apparent side effects.Although these successful examples are still rare and sporadic in the field,they provide the proof of concept for harnessing the power of gene therapy to treat genetic diseases and to modernize our medication.In addition,those success stories illuminate the path for the development of gene therapy treating other genetic diseases.Because of the differences in target organs and cells,distinct barriers to gene delivery exist in gene therapy for each genetic disease.It is not feasible for authors to review the current development in the entire field.Thus,in this article,we will focus onwhatwe can learn from the current success in gene therapy for retinal degenerative diseases to speed up the gene therapy development for lung diseases,such as cystic fibrosis.展开更多
There have been significant advancements in the field of retinal gene therapy in the past several years.In particular,therapeutic efficacy has been achieved in three separate human clinical trials conducted to assess ...There have been significant advancements in the field of retinal gene therapy in the past several years.In particular,therapeutic efficacy has been achieved in three separate human clinical trials conducted to assess the ability of adeno-associated viruses(AAV)to treat of a type of Leber’s congenital amaurosis caused by RPE65 mutations.However,despite the success of retinal gene therapy with AAV,challenges remain for delivering large therapeutic genes or genes requiring long DNA regulatory elements for controlling their expression.For example,Stargardt’s disease,a form of juvenile macular degeneration,is caused by defects in ABCA4,a gene that is too large to be packaged in AAV.Therefore,we investigated the ability of helper dependent adenovirus(HD-Ad)to deliver genes to the retina as it has a much larger transgene capacity.Using an EGFP reporter,our results showed that HD-Ad can transduce the entire retinal epithelium of a mouse using a dose of only 1105 infectious units and maintain transgene expression for at least 4 months.The results demonstrate that HD-Ad has the potential to be an effective vector for the gene therapy of the retina.展开更多
Reproducibility has always been a serious challenge when medical researchers in both academia and industry have tried to build upon previously published discoveries.Blindly chasing faulty results has incurred a huge w...Reproducibility has always been a serious challenge when medical researchers in both academia and industry have tried to build upon previously published discoveries.Blindly chasing faulty results has incurred a huge waste of human and monetary resources.The damage to the progress of scientific discoveries,as well as their application to human well-being,cannot be overestimated.According to two reports by Bayer and Amgen published in 2011 and 2012,64e89%of the socalled“landmark”results could not be reproduced in their pre-clinical validation experiments.1,2 One plausible explanation for this out of proportion irreproducibility is related to the intricacy of the scientific experiments,including the sourcing of reagent antibodies and cell lines,which are major sources of variations.To make validation meaningful,the study materials used in the original studies need to be authenticated so that variations due to the faulty materials can be prevented during follow-up studies.However,the technical complexity and the costs of authentication often discourage this practice in research laboratories.展开更多
The majority of clinical blindness is caused by a loss of transparency of the lens and cornea,largely due to cataracts and corneal injuries.The most common treatment used to restore the transparency is surgical remova...The majority of clinical blindness is caused by a loss of transparency of the lens and cornea,largely due to cataracts and corneal injuries.The most common treatment used to restore the transparency is surgical removal of the damaged tissues,followed by transplantation of donated corneal tissue or an artificial lens.However,these therapies are not without limitations or untoward effects.Unraveling the intricate regulatory signals required for cornea and lens development has made it possible to harness the lineage growth potential of stem cells for cornea repair and lens regeneration,as showcased in two recent studies published in the March 17th issue of Nature.展开更多
Induced pluripotent stem cells(iPSCs)hold great promise for the treatment of human diseases.Two recent first-of-its-kind clinical case reports on the iPSC-based treatment of age-related macular degeneration(AMD)highli...Induced pluripotent stem cells(iPSCs)hold great promise for the treatment of human diseases.Two recent first-of-its-kind clinical case reports on the iPSC-based treatment of age-related macular degeneration(AMD)highlight the hopes and challenges associated with the clinical application of iPSCs.展开更多
Duchenne muscular dystrophy(DMD)is a progressive muscle degenerative disease affecting one out of 3500 male births.Patients usually succumb to the disease by age 25.It has been shown that skipping exons of the DMD gen...Duchenne muscular dystrophy(DMD)is a progressive muscle degenerative disease affecting one out of 3500 male births.Patients usually succumb to the disease by age 25.It has been shown that skipping exons of the DMD gene that contain disease-causing mutations from the pre-mRNA can result in a shortened,but functional,dystrophin protein that could bring clinical benefits to patients.A recent breakthrough has been reported in Science by three groups who demonstrated that genetically deleting exon 23 by gene editing can restore the expression of dystrophin(albeit a shortened version)and improve the muscle function in a mouse model of DMD.展开更多
We devote this short piece to highlight one recent article published in Cell Stem Cell,reporting the correction of large chromosomal inversions of the factor VIII(F8)gene in cells from Hemophilia A patients using the ...We devote this short piece to highlight one recent article published in Cell Stem Cell,reporting the correction of large chromosomal inversions of the factor VIII(F8)gene in cells from Hemophilia A patients using the CRISPR-Cas9 technology,one of the first attempts to edit large segments of chromosomes in patient cells using such methodology.The corrected cells were found free of off-target mutations and producing functional factor VIII in hemophilia mouse model.This work heralds another major advance in bringing CRISPR closer to the therapeutic reality.展开更多
The advent of induced pluripotent stem cells(iPSCs)marked a giant step forward towards the reality of converting one type of primary somatic cells into different lineages capable of clinically repairing damaged tissue...The advent of induced pluripotent stem cells(iPSCs)marked a giant step forward towards the reality of converting one type of primary somatic cells into different lineages capable of clinically repairing damaged tissues and organs.However,the major drawbacks of iPSCs hinder their quick translation to the bedside.These drawbacks include the time-,cost-,and labor-intensive process in production of clinical products from iPSCs,and the inherent risk of long-term tumorigenesis due to the forced expression of transcription factors associated with pluripotency,which are often implicated as aberrations within the cancerous gene circuitry.展开更多
基金supported by grants from RG is an awardee of The Chinese Scholarship Council(CSC)partially supported by grants from The Canadian Institutes of Health Research to JH+1 种基金The National Basic Research Program of China(2011CB9107040)The National Natural Science Foundation of China(30973289,81272971)~~
文摘The role of Ese-2, an Ets family transcription factor, in gene regulation is not known. In this study, the interactionbetween Ese-2 and cytokeratin 18 (K18) intron 1 was characterized in lung epithelial cells. Reporter gene assays showedEse-2 was able to upregulate K18 intron 1 enhanced reporter gene expression by approximately 2-fold. We found thatfull length Ese-2 did not bind DNA strongly, therefore truncated versions of the protein, containing the ETS domain orPointed domain, were created and tested in electrophoresis mobility shift assays. Multiple interactions between the ETSdomain and putative DNA binding sites within K18 intron 1 were observed, which led to the determination of a possibleEse-2 DNA binding consensus sequence. These experiments suggest that Ese-2 could play a role in the regulation ofK18 expression in lung epithelial cells.
基金Work in the authors’laboratories was supported in part by research grants from the National Institutes of Health(AT004418 to TCH)from the Canadian Institutes of Health Research(MOP 125882 to JH).
文摘Vision loss or impairment resulting from the degeneration of the retinal pigment epithelium and photoreceptor death affects millions worldwide.Recent exciting results from clinical studies of small numbers of patients treated with human embryonic stem cell-derived retinal pigment epithelial cells may provide hope for affected individuals.
文摘Asthma is a chronic lung disease that primarily affects the lower respiratory tract in more than 300 million people worldwide.Asthma is often triggered by exposure to certain substances or conditions,such as allergens or cold air.The overreaction to the allergens causes inflammation and narrowing of the airway,which leads to episodic or persistent symptoms including shortness of breath,nocturnal cough,chest tightness,and wheezing.
基金Research in our laboratories was supported by Operating Grants from the Canadian Institutes of Health Research,the Canadian Cystic Fibrosis Foundation,and the Foundation Fighting Blindness-Canada.
文摘After two decades of ups and downs,gene therapy has recently achieved a milestone in treating patients with Leber’s congenital amaurosis(LCA).LCA is a group of inherited blinding diseases with retinal degeneration and severe vision loss in early infancy.Mutations in several genes,including RPE65,cause the disease.Using adenoassociated virus as a vector,three independent teams of investigators have recently shown that RPE65 can be delivered to retinal pigment epithelial cells of LCA patients by subretinal injections resulting in clinical benefits without side effects.However,considering the whole field of gene therapy,there are still major obstacles to clinical applications for other diseases.These obstacles include innate and immune barriers to vector delivery,toxicity of vectors and the lack of sustained therapeutic gene expression.Therefore,new strategies are needed to overcome these hurdles for achieving safe and effective gene therapy.In this article,we shall review the major advancements over the past two decades and,using lung gene therapy as an example,discuss the current obstacles and possible solutions to provide a roadmap for future gene therapy research.
文摘Innate immune responses form the first line of defense against foreign insults and recently significant advances have been made in our understanding of the initiation of innate immune response along with its ability to modulate inflammation. In airway diseases such as asthma, COPD and cystic fibrosis, over reacting of the airway innate immune responses leads to cytokine imbalance and airway remodeling or damage. Helper-dependent adenoviral vectors have the potential to deliver genes to modulate airway innate immune responses and have many advantages over its predecessors. However, there still are a few limitations that need to be addressed prior to their use in clinical applications.
文摘Gene therapy has been considered as the most ideal medical intervention for genetic diseases because it is intended to target the cause of diseases instead of disease symptoms.Availability of techniques for identification of genetic mutations and for in vitro manipulation of genes makes it practical and attractive.After the initial hype in 1990s and later disappointments in clinical trials formore than a decade,light has finally come into the tunnel in recent years,especially in the field of eye gene therapy where it has taken big strides.Clinical trials in gene therapy for retinal degenerative diseases such as Leber’s congenital amaurosis(LCA)and choroideremia demonstrated clear therapeutic efficacies without apparent side effects.Although these successful examples are still rare and sporadic in the field,they provide the proof of concept for harnessing the power of gene therapy to treat genetic diseases and to modernize our medication.In addition,those success stories illuminate the path for the development of gene therapy treating other genetic diseases.Because of the differences in target organs and cells,distinct barriers to gene delivery exist in gene therapy for each genetic disease.It is not feasible for authors to review the current development in the entire field.Thus,in this article,we will focus onwhatwe can learn from the current success in gene therapy for retinal degenerative diseases to speed up the gene therapy development for lung diseases,such as cystic fibrosis.
文摘There have been significant advancements in the field of retinal gene therapy in the past several years.In particular,therapeutic efficacy has been achieved in three separate human clinical trials conducted to assess the ability of adeno-associated viruses(AAV)to treat of a type of Leber’s congenital amaurosis caused by RPE65 mutations.However,despite the success of retinal gene therapy with AAV,challenges remain for delivering large therapeutic genes or genes requiring long DNA regulatory elements for controlling their expression.For example,Stargardt’s disease,a form of juvenile macular degeneration,is caused by defects in ABCA4,a gene that is too large to be packaged in AAV.Therefore,we investigated the ability of helper dependent adenovirus(HD-Ad)to deliver genes to the retina as it has a much larger transgene capacity.Using an EGFP reporter,our results showed that HD-Ad can transduce the entire retinal epithelium of a mouse using a dose of only 1105 infectious units and maintain transgene expression for at least 4 months.The results demonstrate that HD-Ad has the potential to be an effective vector for the gene therapy of the retina.
基金Work in the authors’laboratories was supported in part by research grants from the National Institutes of Health(CA172233 to KP,AT004418 to TCH)Canadian Institutes of Health Research(MOP 125882 to JH).
文摘Reproducibility has always been a serious challenge when medical researchers in both academia and industry have tried to build upon previously published discoveries.Blindly chasing faulty results has incurred a huge waste of human and monetary resources.The damage to the progress of scientific discoveries,as well as their application to human well-being,cannot be overestimated.According to two reports by Bayer and Amgen published in 2011 and 2012,64e89%of the socalled“landmark”results could not be reproduced in their pre-clinical validation experiments.1,2 One plausible explanation for this out of proportion irreproducibility is related to the intricacy of the scientific experiments,including the sourcing of reagent antibodies and cell lines,which are major sources of variations.To make validation meaningful,the study materials used in the original studies need to be authenticated so that variations due to the faulty materials can be prevented during follow-up studies.However,the technical complexity and the costs of authentication often discourage this practice in research laboratories.
基金Work in Dr.Jim Hu’s laboratory was supported in part by research grants from the Canadian Institutes of Health Research(MOP 125882)Cystic Fibrosis Canada(Grant ID#3023)Cystic Fibrosis Foundation Therapeutics(HU15XX0).
文摘The majority of clinical blindness is caused by a loss of transparency of the lens and cornea,largely due to cataracts and corneal injuries.The most common treatment used to restore the transparency is surgical removal of the damaged tissues,followed by transplantation of donated corneal tissue or an artificial lens.However,these therapies are not without limitations or untoward effects.Unraveling the intricate regulatory signals required for cornea and lens development has made it possible to harness the lineage growth potential of stem cells for cornea repair and lens regeneration,as showcased in two recent studies published in the March 17th issue of Nature.
基金Work in the authors’laboratories was supported in part by research grants from the National Institutes of Health(AT004418 to TCH)from the Canadian Institutes of Health Research(MOP 125882 to JH)+1 种基金a Cystic Fibrosis Foundation Therapeutics,Inc.grant(HU15XX0 to JH)a Cystic Fibrosis Canada grant(#3032 to JH).
文摘Induced pluripotent stem cells(iPSCs)hold great promise for the treatment of human diseases.Two recent first-of-its-kind clinical case reports on the iPSC-based treatment of age-related macular degeneration(AMD)highlight the hopes and challenges associated with the clinical application of iPSCs.
基金Work in the authors’laboratories was supported in part by the Canadian Institutes of Health Research(MOP 125882 to JH)Cystic Fibrosis Canada(Grant ID#3023 to JH)+1 种基金Cystic Fibrosis Foundation Therapeutics(HU15XX0 to JH)research grants from the National Institutes of Health(NS079568,NS082536,DK090380 to XX).LY is a recipient of the Hospital for Sick Children RESTRACOMP studentship.
文摘Duchenne muscular dystrophy(DMD)is a progressive muscle degenerative disease affecting one out of 3500 male births.Patients usually succumb to the disease by age 25.It has been shown that skipping exons of the DMD gene that contain disease-causing mutations from the pre-mRNA can result in a shortened,but functional,dystrophin protein that could bring clinical benefits to patients.A recent breakthrough has been reported in Science by three groups who demonstrated that genetically deleting exon 23 by gene editing can restore the expression of dystrophin(albeit a shortened version)and improve the muscle function in a mouse model of DMD.
基金Work in Dr.Jim Hu’s laboratory was supported in part by a research grant from the Canadian Institutes of Health Research(MOP 125882).
文摘We devote this short piece to highlight one recent article published in Cell Stem Cell,reporting the correction of large chromosomal inversions of the factor VIII(F8)gene in cells from Hemophilia A patients using the CRISPR-Cas9 technology,one of the first attempts to edit large segments of chromosomes in patient cells using such methodology.The corrected cells were found free of off-target mutations and producing functional factor VIII in hemophilia mouse model.This work heralds another major advance in bringing CRISPR closer to the therapeutic reality.
基金Work in Dr.Jim Hu’s laboratory was supported in part by a research grant from the Canadian Institutes of Health Research(MOP 125882).
文摘The advent of induced pluripotent stem cells(iPSCs)marked a giant step forward towards the reality of converting one type of primary somatic cells into different lineages capable of clinically repairing damaged tissues and organs.However,the major drawbacks of iPSCs hinder their quick translation to the bedside.These drawbacks include the time-,cost-,and labor-intensive process in production of clinical products from iPSCs,and the inherent risk of long-term tumorigenesis due to the forced expression of transcription factors associated with pluripotency,which are often implicated as aberrations within the cancerous gene circuitry.
