Imaging structural and functional relationships between cells and ECM

该项目刚刚基于成骨不全（OI）小鼠模型的lunag病理学研究启动。通过将ECM的组织学和免疫荧光成像与同一组织切片中信使RNA（mRNA-FISH）的荧光原位杂交相结合，我们能够鉴定新生儿肺病理（在纯合子动物中致死）是由胎儿呼吸运动期间肺膨胀不足引起的。我们现在正致力于扩展研究，以包括共焦拉曼光谱成像，用于量化相同切片中纤维中胶原蛋白的量和组织（因为肺膨胀不足似乎是由ECM中胶原蛋白沉积不足引起的）。此外，我们正在努力将这些成像模式与基于10 X Genomics的新Xenium检测的定量空间转录组成像以及活E18.5胚胎中肺部的超声背向散射显微镜相结合。 新生儿急性呼吸窘迫是OI和其他骨骼发育不良的共同特征，是这些疾病中新生儿致死的主要原因。传统上，它被归因于出生后影响婴儿的肋骨骨折和畸形，而不是与出生前胎儿呼吸运动不足相关的肺发育不全。在发现新生小鼠严重的肺发育不全后，我们意识到胶原蛋白缺乏和肋骨畸形和骨折可能对子宫内的肺有更明显的影响。如果得到证实，这些发现可能会极大地影响骨骼发育不良新生儿的治疗，提示应预期肺发育不良，立即由新生儿肺病学家进行评估，并进行治疗以提高生存可能性并减少以后的继发性肺部病理学。 在该项目中，通过结合组织切片的组织学、光谱学、RNA和功能成像所揭示的丰富信息表明了进一步推进和开发用于这种多模态成像的技术和方法的重要性。该项目仅在一周前启动。虽然他还没有结果和出版物报告，但我们预计该项目将对我们和其他实验室的结缔组织病理学研究产生重大影响，类似于促使其启动的肺部发现。

This project has just been initiated based on studies of lunag pathology in a mouse model of osteogenesis imperfecta (OI). By combining histological and immunofluorescence imaging of ECM with fluorescence in-situ hybridization of messanger RNA (mRNA-FISH) in the same tissue section we were able to identify that neonatal lung pathology (lethal in homozygous animals) is caused by deficient lung inflation during fetal breathing movements. We are now working on expanding the study to include confocal Raman spectroscopic imaging for quantifying the amount and organization of collagen in fibers in the same sections (since the deficient lung inflation appears to be caused deficient collagen deposition in the ECM). Additionally we are working on integration of these imaging modalities with quantitative spatial transcriptomic imaging based on the new Xenium assay from 10X Genomics and with ultrasound backscatter microscopy of lungs in live E18.5 embryos. Acute respitary distress in neonates is a common feature of OI and other skeletal dysplasias, which is primarily responsible for neonatal lethality in these disorders. Traditionally, it has been attributed to rib cage fractures and deformities affecting the babies after birth but not to lung hypoplasia associated with deficient fetal breathing movements before the birth. Having discovered severe lung hypoplasia in newborn mice, we realized that collagen deficiency and rib cage deformities and fractures may have even more pronounced effect on lungs in utero. If confirmed, these findings may dramatically affect the treatment of newborns with skeletal dysplasias by suggesting that lung hypoplasia should be expected, immediately evaluated by neonatal pulmonologists, and treated to improve the survival likelyhood and reduce secondary lung pathology later in life. The wealth of information revealed by combining histological, spectroscopic, RNA, and functional imaging of tissue sections in this project suggested the importance of further advancing and developing the technology and methods for such multimodal imaging. This project has been initiated only a week ago. While he wave no results and publications to report yet, we anticipate that the project will have a major impact on studies of connective tissue pathology in our and other laborotories similar to the lung findings that prompted its initiation.