Masci Marco, Wang Min, Imbert Laurianne, Barnes Aileen M, Spevak Lyudmila, Lukashova Lyudmila, Huang Yihe, Ma Yan, Marini Joan C, Jacobsen Christina M, Warman Matthew L, Boskey Adele L
Weill Cornell Medical College, New York, NY, United States.
Mineralized Tissues Laboratory, Hospital for Special Surgery, New York, NY, United States.
Bone. 2016 Jun;87:120-9. doi: 10.1016/j.bone.2016.04.011. Epub 2016 Apr 13.
The Col1a2(+/G610C) knock-in mouse, models osteogenesis imperfecta in a large old order Amish family (OOA) with type IV OI, caused by a G-to-T transversion at nucleotide 2098, which alters the gly-610 codon in the triple-helical domain of the α2(I) chain of type I collagen. Mineral and matrix properties of the long bones and vertebrae of male Col1a2(+/G610C) and their wild-type controls (Col1a2(+/+)), were characterized to gain insight into the role of α2-chain collagen mutations in mineralization. Additionally, we examined the rescuability of the composition by sclerostin inhibition initiated by crossing Col1a2(+/G610C) with an LRP(+/A214V) high bone mass allele. At age 10-days, vertebrae and tibia showed few alterations by micro-CT or Fourier transform infrared imaging (FTIRI). At 2-months-of-age, Col1a2(+/G610C) tibias had 13% fewer secondary trabeculae than Col1a2(+/+), these were thinner (11%) and more widely spaced (20%) than those of Col1a2(+/+) mice. Vertebrae of Col1a2(+/G610C) mice at 2-months also had lower bone volume fraction (38%), trabecular number (13%), thickness (13%) and connectivity density (32%) compared to Col1(a2+/+). The cortical bone of Col1a2(+/G610C) tibias at 2-months had 3% higher tissue mineral density compared to Col1a2(+/+); Col1a2(+/G610C) vertebrae had lower cortical thickness (29%), bone area (37%) and polar moment of inertia (38%) relative to Col1a2(+/+). FTIRI analysis, which provides information on bone chemical composition at ~7μm-spatial resolution, showed tibias at 10-days did not differ between genotypes. Comparing identical bone types in Col1a2(+/G610C) to Col1a2(+/+) at 2-months-of-age, tibias showed higher mineral-to-matrix ratio in trabeculae (17%) and cortices (31%). and in vertebral cortices (28%). Collagen maturity was 42% higher at 10-days-of-age in Col1a2(+/G610C) vertebral trabeculae and in 2-month tibial cortices (12%), vertebral trabeculae (42%) and vertebral cortices (12%). Higher acid-phosphate substitution was noted in 10-day-old trabecular bone in vertebrae (31%) and in 2-month old trabecular bone in both tibia (31%) and vertebrae (4%). There was also a 16% lower carbonate-to-phosphate ratio in vertebral trabeculae and a correspondingly higher (22%) carbonate-to-phosphate ratio in 2month-old vertebral cortices. At age 3-months-of-age, male femurs with both a Col1a2(+/G610C) allele and a Lrp5 high bone mass allele (Lrp5+/A214V) showed an improvement in bone composition, presenting higher trabecular carbonate-to-phosphate ratio (18%) and lower trabecular and cortical acid-phosphate substitutions (8% and 18%, respectively). Together, these results indicate that mutant collagen α2(I) chain affects both bone quantity and composition, and the usefulness of this model for studies of potential OI therapies such as anti-sclerostin treatments.
Col1a2(+/G610C)基因敲入小鼠模拟了一个大型老派阿米什家族(OOA)中的IV型成骨不全症,该病症由核苷酸2098处的G到T颠换引起,这改变了I型胶原蛋白α2(I)链三螺旋结构域中的gly-610密码子。对雄性Col1a2(+/G610C)及其野生型对照(Col1a2(+/+))的长骨和椎骨的矿物质和基质特性进行了表征,以深入了解α2链胶原蛋白突变在矿化中的作用。此外,我们通过将Col1a2(+/G610C)与LRP(+/A214V)高骨量等位基因杂交引发的硬化蛋白抑制作用,研究了其对骨成分的挽救能力。在10日龄时,通过微型计算机断层扫描(micro-CT)或傅里叶变换红外成像(FTIRI)观察到椎骨和胫骨几乎没有变化。在2月龄时,Col1a2(+/G610C)胫骨的次级小梁比Col1a2(+/+)少13%,这些小梁比Col1a2(+/+)小鼠的小梁更薄(11%)且间距更宽(20%)。2月龄Col1a2(+/G610C)小鼠的椎骨与Col1(a2+/+)相比,骨体积分数也更低(38%)、小梁数量(13%)、厚度(13%)和连接密度(32%)。2月龄Col1a2(+/G610C)胫骨的皮质骨组织矿物质密度比Col1a2(+/+)高3%;与Col1a2(+/+)相比,Col1a2(+/G610C)椎骨的皮质厚度更低(29%)、骨面积更低(37%)和极惯性矩更低(38%)。FTIRI分析可在约7μm的空间分辨率下提供骨化学成分信息,结果显示10日龄时不同基因型的胫骨之间没有差异。比较2月龄时Col1a2(+/G610C)与Col1a2(+/+)相同类型的骨,胫骨的小梁(17%)、皮质(31%)以及椎骨皮质(28%)中的矿物质与基质比率更高。在10日龄时,Col1a2(+/G610C)椎骨小梁以及2月龄胫骨皮质(12%)、椎骨小梁(42%)和椎骨皮质(12%)中的胶原蛋白成熟度高42%。在10日龄的椎骨小梁(31%)以及2月龄的胫骨(31%)和椎骨(4%)小梁骨中观察到更高的酸性磷酸盐取代。椎骨小梁中的碳酸盐与磷酸盐比率也降低了16%,相应地,2月龄椎骨皮质中的碳酸盐与磷酸盐比率更高(22%)。在3月龄时,同时携带Col1a2(+/G610C)等位基因和Lrp5高骨量等位基因(Lrp5+/A214V)的雄性股骨的骨成分有所改善,呈现出更高的小梁碳酸盐与磷酸盐比率(18%)以及更低的小梁和皮质酸性磷酸盐取代(分别为8%和18%)。总之,这些结果表明突变的胶原蛋白α2(I)链会影响骨量和骨成分,并且该模型对于研究潜在的成骨不全症治疗方法(如抗硬化蛋白治疗)具有实用性。