KIBRA基因多态性与默认网络、执行控制网络相关性的多模态MRI研究
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摘要
目的：
基因学研究表明KIBRA rs17070145位点多态性与人类情景记忆、执行功能以及阿尔兹海默病密切相关,而其神经机制尚不清楚。本研究采用多模态磁共振成像(magnetic resonance imaging, MRI)技术,联合运用基于体素的形态学(voxel-based morphometry analysis, VBM)-.独立成分分析(independent component analysis, ICA)方法,研究KIBRA基因多态性与大脑默认网络、执行控制网络的关系。
材料与方法：
选择符合入组标准的健康青年志愿者288例,平均年龄：22.8±2.5岁；男性133例,女性155例。每名受试者完成韦氏记忆量表、威斯康星卡片测评。利用GE3.0T Signa HDX磁共振扫描仪对所有受试者进行静息态、结构像以及弥散像数据的采集。静息态扫描时嘱受试者闭眼、保持身体静止不动,均匀呼吸,精神放松,尽量不思考任何事情。
1.基因型测定,利用EZgeneTM血液gDNA小量提取试剂盒,从3000μl全血中提取受试者的基因组DNA,应用聚合酶链反应(polymerase chain reaction, PCR)-连接酶检测反应(ligation detection reaction, LDR)的方法,检测受试者KIBRA基因rs17070145位点C/T等位基因多态性；根据不同基因型将受试者分成两组,一组为C等位基因携带者,另一组为TT纯合子携带者。
2.采用基于Matlab平台的SPM8软件及其插件包VBM8对高分辨率结构像进行预处理,预处理过程包括：灰、白质分割、空间标准化及空间平滑。采用基于Matlab平台的DPARSF软件对静息态功能数据进行预处理,预处理过程包括：时间校正、头动校正、空间标准化、重采样到2mm×2mm×2mm的立方体素及空间平滑。
3.采用预处理后的静息态功能数据,利用基于Matlab平台的MICA软件进行组ICA分析,得到5个静息态脑网络,在控制年龄、性别以及受教育年限影响因素后,探讨KIBRA基因对大脑网络功能的影响。
4.采用预处理后的高分辨率结构像以及弥散像数据,在控制年龄、性别以及受教育年限影响因素后,研究KIBRA基因对大脑灰质体积、白质体积、白质完整性的影响。
5.并采用AlphSim方法对结果进行多重比较校正,最后将校正后的统计参数映射到MNI标准三维模板脑进行显示,观察与流体推理有显著相关性的区域。描述并记录有统计学意义脑区的团块位置、大小(cluster size)、MNI坐标及相关强度。
结果：
1.通过KIBRA基因分型检测,288例受试者中,121例为C等位基因携带者(包括14例CC纯合子携带者),167例TT纯合子携带者；基因型分布频率符合Hardy-Weinberg分布[χ2(2)=0.36,p=0.55]；年龄、性别、受教育年限、韦氏记忆量表评分以及威斯康辛卡片评分未发现组间差异,但CC纯合子携带者的记忆、执行功能有降低的趋势。
2.通过组ICA方法,获得5个经典的静息态脑网络：默认网络(前、后两部分)、执行控制网络、感觉运动网络、视觉网络。
3.与KIBRA TT等位基因携带者相比,C等位基因携带者在默认网络的内侧前额叶皮层、后扣带皮层,以及执行控制网络的右侧岛叶、双侧尾状核、背侧前扣带皮层的脑同步性活动明显增加；而视觉网络、感觉运动网络却没有发现明显差异。
4. KIBRA C等位基因携带者在默认网络的内侧前额叶、执行控制网络的双侧的背侧前扣带皮层灰质体积较TT等位基因携带者明显减少。大脑白质体积、白质FA值(fractional anisotropy, FA)均未发现组间差异。
结论：
1.默认网络、执行控制网络的结构及功能受到KIBRA基因多态性的影响；
2.在正常年轻受试者中,KIBRA基因C携带者脑网络同步性活动的增加则可能是对脑局部灰质体积减少的一种代偿；
3.在正常年轻受试者中,KIBRA基因多态性可能不会影响大脑白质的完整性。
Objective:
Genetic variation at the KIBRA rs17070145polymorphism has been linked to episodic memory, executive function, and Alzheimer's disease (AD); however its neural substrates remain elusive. We combined voxel-based morphometry (VBM) analysis of structural MRI data and independent component analysis (ICA) analysis of resting-state fMRI data to investigate to investigate neural correlates of KIBRA gene and brain networks in a large sample of healthy young adults.
Subjects and Methods:
A total of288healthy, young, right-handed subjects were recruited in this study. Resting-state fMRI, high-resolution structural imaging, and diffusion tensor imaging were performed using a GE3.0T Signa HDX scanner. During the scanning, all subjects were explicitly instructed to keep their eyes closed, relax, as motionless as possible and think of nothing.
1. Genotyping We extracted genomic DNA from3000μl of whole blood using the EZgeneTM Blood gDNA Miniprep Kit (Biomiga Inc, San Diego, CA, USA). Then, we genotyped the KIBRA rs17070145in each subject using the PCR and ligation detection reaction (LDR) method.
2. All preprocessing steps were carried out using the statistical parametric mapping (SPM8, http://www.fil.ion.ucl.ac.uk/spm) based on Matlab.
3. The preprocessing of high resolution structural images including segmentation of gray and white matter, spatial normalization and smooth with a4mm full width at half maximum; while the preprocessing of rs-fMRI data including slice timing, realignment, spatial normalization to the MNI, resampling to to2×2×2mm3cubic voxels, and smoothed with a4mm full width at half maximum.
4. Group ICA was performed using the MICA software based on Matlab, five RSNs were chosen from the results of ICA; and the group sychroniztion differences were investigated between the two KIBRA genotypes with the age, gender and education years as covariates.
5. Voxel-based morphometry (VBM) analysis of structural MRI data was performed to investigate group differences of grey matter volume and white matter integrity
6. Multiple comparisons were statistically corrected by Monte Carlo simulation. Finally, the corrected statistical parameter mapping was overlapped onto MNI standard template and the coordinate, size and peak t values of each significant cluster were recorded.
Results:
1. The genotype distribution of the SNP was in Hardy-Weinberg equilibrium [χ2(2)=0.36, p-0.55]. There were no significant group differences in age, gender, years of education, memory and WCST scores although the CC group showed a trend towards decreased memory and executive function.
2. Five meaningful resting state networks were obtained from the MICA results: default mode network (anterior, posterior), executive control network, sensorimotor netwok, and visual network.
3. KIBRA C-allele carriers showed significantly increased synchronization in the right MPFC of the aDMN and left PCC of the pDMN, the right AI, bilateral Cau, and bilateral dACC of ECN. No significant differences in synchronization were found in either the SMN or the VN between the two genotypes.
4. KIBRA C-allele carriers showed significant decrease GMV in the right MPFC of the aDMN and the dACC of the ECN. No significant differences in white matter volume and white matter integrity were found between the two genotypes.
Conclusion:
1. The structure and function of the DMN and ECN may be selectively affected by KIBRA rs17070145polymorphism;
2. The increased synchronization in the DMN and ECN is likely a reflection of compensation for the GMV deficit in these networks in young healthy subjects.
3. The white matter volume and integrity might not be affected by KIBRA rs17070145polymorphism.
基因学研究表明KIBRA rs17070145位点多态性与人类情景记忆、执行功能以及阿尔兹海默病密切相关,而其神经机制尚不清楚。本研究采用多模态磁共振成像(magnetic resonance imaging, MRI)技术,联合运用基于体素的形态学(voxel-based morphometry analysis, VBM)-.独立成分分析(independent component analysis, ICA)方法,研究KIBRA基因多态性与大脑默认网络、执行控制网络的关系。
材料与方法：
选择符合入组标准的健康青年志愿者288例,平均年龄：22.8±2.5岁；男性133例,女性155例。每名受试者完成韦氏记忆量表、威斯康星卡片测评。利用GE3.0T Signa HDX磁共振扫描仪对所有受试者进行静息态、结构像以及弥散像数据的采集。静息态扫描时嘱受试者闭眼、保持身体静止不动,均匀呼吸,精神放松,尽量不思考任何事情。
1.基因型测定,利用EZgeneTM血液gDNA小量提取试剂盒,从3000μl全血中提取受试者的基因组DNA,应用聚合酶链反应(polymerase chain reaction, PCR)-连接酶检测反应(ligation detection reaction, LDR)的方法,检测受试者KIBRA基因rs17070145位点C/T等位基因多态性；根据不同基因型将受试者分成两组,一组为C等位基因携带者,另一组为TT纯合子携带者。
2.采用基于Matlab平台的SPM8软件及其插件包VBM8对高分辨率结构像进行预处理,预处理过程包括：灰、白质分割、空间标准化及空间平滑。采用基于Matlab平台的DPARSF软件对静息态功能数据进行预处理,预处理过程包括：时间校正、头动校正、空间标准化、重采样到2mm×2mm×2mm的立方体素及空间平滑。
3.采用预处理后的静息态功能数据,利用基于Matlab平台的MICA软件进行组ICA分析,得到5个静息态脑网络,在控制年龄、性别以及受教育年限影响因素后,探讨KIBRA基因对大脑网络功能的影响。
4.采用预处理后的高分辨率结构像以及弥散像数据,在控制年龄、性别以及受教育年限影响因素后,研究KIBRA基因对大脑灰质体积、白质体积、白质完整性的影响。
5.并采用AlphSim方法对结果进行多重比较校正,最后将校正后的统计参数映射到MNI标准三维模板脑进行显示,观察与流体推理有显著相关性的区域。描述并记录有统计学意义脑区的团块位置、大小(cluster size)、MNI坐标及相关强度。
结果：
1.通过KIBRA基因分型检测,288例受试者中,121例为C等位基因携带者(包括14例CC纯合子携带者),167例TT纯合子携带者；基因型分布频率符合Hardy-Weinberg分布[χ2(2)=0.36,p=0.55]；年龄、性别、受教育年限、韦氏记忆量表评分以及威斯康辛卡片评分未发现组间差异,但CC纯合子携带者的记忆、执行功能有降低的趋势。
2.通过组ICA方法,获得5个经典的静息态脑网络：默认网络(前、后两部分)、执行控制网络、感觉运动网络、视觉网络。
3.与KIBRA TT等位基因携带者相比,C等位基因携带者在默认网络的内侧前额叶皮层、后扣带皮层,以及执行控制网络的右侧岛叶、双侧尾状核、背侧前扣带皮层的脑同步性活动明显增加；而视觉网络、感觉运动网络却没有发现明显差异。
4. KIBRA C等位基因携带者在默认网络的内侧前额叶、执行控制网络的双侧的背侧前扣带皮层灰质体积较TT等位基因携带者明显减少。大脑白质体积、白质FA值(fractional anisotropy, FA)均未发现组间差异。
结论：
1.默认网络、执行控制网络的结构及功能受到KIBRA基因多态性的影响；
2.在正常年轻受试者中,KIBRA基因C携带者脑网络同步性活动的增加则可能是对脑局部灰质体积减少的一种代偿；
3.在正常年轻受试者中,KIBRA基因多态性可能不会影响大脑白质的完整性。
Objective:
Genetic variation at the KIBRA rs17070145polymorphism has been linked to episodic memory, executive function, and Alzheimer's disease (AD); however its neural substrates remain elusive. We combined voxel-based morphometry (VBM) analysis of structural MRI data and independent component analysis (ICA) analysis of resting-state fMRI data to investigate to investigate neural correlates of KIBRA gene and brain networks in a large sample of healthy young adults.
Subjects and Methods:
A total of288healthy, young, right-handed subjects were recruited in this study. Resting-state fMRI, high-resolution structural imaging, and diffusion tensor imaging were performed using a GE3.0T Signa HDX scanner. During the scanning, all subjects were explicitly instructed to keep their eyes closed, relax, as motionless as possible and think of nothing.
1. Genotyping We extracted genomic DNA from3000μl of whole blood using the EZgeneTM Blood gDNA Miniprep Kit (Biomiga Inc, San Diego, CA, USA). Then, we genotyped the KIBRA rs17070145in each subject using the PCR and ligation detection reaction (LDR) method.
2. All preprocessing steps were carried out using the statistical parametric mapping (SPM8, http://www.fil.ion.ucl.ac.uk/spm) based on Matlab.
3. The preprocessing of high resolution structural images including segmentation of gray and white matter, spatial normalization and smooth with a4mm full width at half maximum; while the preprocessing of rs-fMRI data including slice timing, realignment, spatial normalization to the MNI, resampling to to2×2×2mm3cubic voxels, and smoothed with a4mm full width at half maximum.
4. Group ICA was performed using the MICA software based on Matlab, five RSNs were chosen from the results of ICA; and the group sychroniztion differences were investigated between the two KIBRA genotypes with the age, gender and education years as covariates.
5. Voxel-based morphometry (VBM) analysis of structural MRI data was performed to investigate group differences of grey matter volume and white matter integrity
6. Multiple comparisons were statistically corrected by Monte Carlo simulation. Finally, the corrected statistical parameter mapping was overlapped onto MNI standard template and the coordinate, size and peak t values of each significant cluster were recorded.
Results:
1. The genotype distribution of the SNP was in Hardy-Weinberg equilibrium [χ2(2)=0.36, p-0.55]. There were no significant group differences in age, gender, years of education, memory and WCST scores although the CC group showed a trend towards decreased memory and executive function.
2. Five meaningful resting state networks were obtained from the MICA results: default mode network (anterior, posterior), executive control network, sensorimotor netwok, and visual network.
3. KIBRA C-allele carriers showed significantly increased synchronization in the right MPFC of the aDMN and left PCC of the pDMN, the right AI, bilateral Cau, and bilateral dACC of ECN. No significant differences in synchronization were found in either the SMN or the VN between the two genotypes.
4. KIBRA C-allele carriers showed significant decrease GMV in the right MPFC of the aDMN and the dACC of the ECN. No significant differences in white matter volume and white matter integrity were found between the two genotypes.
Conclusion:
1. The structure and function of the DMN and ECN may be selectively affected by KIBRA rs17070145polymorphism;
2. The increased synchronization in the DMN and ECN is likely a reflection of compensation for the GMV deficit in these networks in young healthy subjects.
3. The white matter volume and integrity might not be affected by KIBRA rs17070145polymorphism.
引文
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