Background:To provide an understanding of important aspects of the participant recruitment and data collec-tion,become aware of any potential problems,and obtain necessary information in order to design a large-scale ...Background:To provide an understanding of important aspects of the participant recruitment and data collec-tion,become aware of any potential problems,and obtain necessary information in order to design a large-scale randomized controlled trial(RCT)for lung cancer and colorectal cancer(CRC)screening in China.Methods:This feasibility study was a multicentered,open-label,pilot randomized trial.A total of 2696 participants who were at high risk of lung cancer were recruited from three screening centers and randomly allocated to arm 1(n=894),annual low-dose computed tomography(LDCT)plus a baseline colonoscopy;arm 2(n=902),biennial LDCT plus annual fecal immunochemical test(FIT)with OC-Sensor(OC-FIT);and arm 3(n=900),annual Insure-FIT plus Septin 9 blood test.Information on randomization,compliance,positivity rate,cancer case detection,and contamination with screening for lung cancer and CRC were collected.Results:Participant characteristics were similar across study arms.The compliance rate of annual LDCT screening in arm 1 was 86.4%(95%CI:83.9%,88.5%)at baseline(T0),and 69.0%(95%CI:65.8%,72.0%)and 70.7%(95%CI:67.6%,73.7%)at the following two rounds(T1 and T2).The compliance rates of biennial LDCT screening in arm 2 were similar to those in arm 1 in the corresponding rounds.The compliance rate was 55.5%(95%CI:52.2%,58.8%)for colonoscopy in arm 1,while the compliance rates of OC-FIT,Insure-FIT,and the Septin 9 test in arms 2 and 3 were all approximately 90%at T0,decreasing to 65%-80%at T1 and T2.The positivity rate,cancer case detection rate,and contamination rate of screening for lung cancer and CRC were also reported.Conclusion:In this pilot study,the feasibility of an RCT in China of lung cancer and CRC screening was demon-strated.展开更多
Liquid biopsy is a highly promising method for non-invasive detection of tumor-associated nucleic acid fragments in body fluids but is challenged by the low abundance of nucleic acids of clinical interest and their se...Liquid biopsy is a highly promising method for non-invasive detection of tumor-associated nucleic acid fragments in body fluids but is challenged by the low abundance of nucleic acids of clinical interest and their sequence homology with the vast background of nucleic acids from healthy cells.Recently,programmable endonucleases such as clustered regularly interspaced short palindromic repeats(CRISPR)associated protein(Cas)and prokaryotic Argonautes have been successfully used to remove background nucleic acids and enrich mutant allele fractions,enabling their detection with deep next generation sequencing(NGS).However,the enrichment level achievable with these assays is limited by futile binding events and off-target cleavage.To overcome these shortcomings,we conceived a new assay(Programmable Enzyme-Assisted Selective Exponential Amplification,PASEA)that combines the cleavage of wild type alleles with concurrent polymerase amplification.While PASEA increases the numbers of both wild type and mutant alleles,the numbers of mutant alleles increase at much greater rates,allowing PASEA to achieve an unprecedented level of selective enrichment of targeted alleles.By combining CRISPR-Cas9 based cleavage with recombinase polymerase amplification,we converted samples with0.01%somatic mutant allele fractions(MAFs)to products with 70%MAFs in a single step within 20 min,enabling inexpensive,rapid genotyping with such as Sanger sequencers.Furthermore,PASEA's extraordinary efficiency facilitates sensitive real-time detection of somatic mutant alleles at the point of care with custom designed Exo-RPA probes.Real-time PASEA'performance was proved equivalent to clinical amplification refractory mutation system(ARMS)-PCR and NGS when testing over hundred cancer patients'samples.This strategy has the potential to reduce the cost and time of cancer screening and genotyping,and to enable targeted therapies in resource-limited settings.展开更多
文摘Background:To provide an understanding of important aspects of the participant recruitment and data collec-tion,become aware of any potential problems,and obtain necessary information in order to design a large-scale randomized controlled trial(RCT)for lung cancer and colorectal cancer(CRC)screening in China.Methods:This feasibility study was a multicentered,open-label,pilot randomized trial.A total of 2696 participants who were at high risk of lung cancer were recruited from three screening centers and randomly allocated to arm 1(n=894),annual low-dose computed tomography(LDCT)plus a baseline colonoscopy;arm 2(n=902),biennial LDCT plus annual fecal immunochemical test(FIT)with OC-Sensor(OC-FIT);and arm 3(n=900),annual Insure-FIT plus Septin 9 blood test.Information on randomization,compliance,positivity rate,cancer case detection,and contamination with screening for lung cancer and CRC were collected.Results:Participant characteristics were similar across study arms.The compliance rate of annual LDCT screening in arm 1 was 86.4%(95%CI:83.9%,88.5%)at baseline(T0),and 69.0%(95%CI:65.8%,72.0%)and 70.7%(95%CI:67.6%,73.7%)at the following two rounds(T1 and T2).The compliance rates of biennial LDCT screening in arm 2 were similar to those in arm 1 in the corresponding rounds.The compliance rate was 55.5%(95%CI:52.2%,58.8%)for colonoscopy in arm 1,while the compliance rates of OC-FIT,Insure-FIT,and the Septin 9 test in arms 2 and 3 were all approximately 90%at T0,decreasing to 65%-80%at T1 and T2.The positivity rate,cancer case detection rate,and contamination rate of screening for lung cancer and CRC were also reported.Conclusion:In this pilot study,the feasibility of an RCT in China of lung cancer and CRC screening was demon-strated.
基金supported by China Scholarship CouncilNIH grant to the University of Pennsylvania(No.K011K01TW011190-01A1)+1 种基金NIH grant to the University of Pennsylvania(No.R21CA228614-01A1)Beijing Hope Run Special Fund from the Cancer Foundation of China(Nos.LC2019L04 and LC2020A36)。
文摘Liquid biopsy is a highly promising method for non-invasive detection of tumor-associated nucleic acid fragments in body fluids but is challenged by the low abundance of nucleic acids of clinical interest and their sequence homology with the vast background of nucleic acids from healthy cells.Recently,programmable endonucleases such as clustered regularly interspaced short palindromic repeats(CRISPR)associated protein(Cas)and prokaryotic Argonautes have been successfully used to remove background nucleic acids and enrich mutant allele fractions,enabling their detection with deep next generation sequencing(NGS).However,the enrichment level achievable with these assays is limited by futile binding events and off-target cleavage.To overcome these shortcomings,we conceived a new assay(Programmable Enzyme-Assisted Selective Exponential Amplification,PASEA)that combines the cleavage of wild type alleles with concurrent polymerase amplification.While PASEA increases the numbers of both wild type and mutant alleles,the numbers of mutant alleles increase at much greater rates,allowing PASEA to achieve an unprecedented level of selective enrichment of targeted alleles.By combining CRISPR-Cas9 based cleavage with recombinase polymerase amplification,we converted samples with0.01%somatic mutant allele fractions(MAFs)to products with 70%MAFs in a single step within 20 min,enabling inexpensive,rapid genotyping with such as Sanger sequencers.Furthermore,PASEA's extraordinary efficiency facilitates sensitive real-time detection of somatic mutant alleles at the point of care with custom designed Exo-RPA probes.Real-time PASEA'performance was proved equivalent to clinical amplification refractory mutation system(ARMS)-PCR and NGS when testing over hundred cancer patients'samples.This strategy has the potential to reduce the cost and time of cancer screening and genotyping,and to enable targeted therapies in resource-limited settings.
