气管内注射人诱导多能干细胞源性的肺泡细胞和内皮细胞在远端肺部定植，并改善大鼠肺气肿模型。

Intratracheally injected human-induced pluripotent stem cell-derived pneumocytes and endothelial cells engraft in the distal lung and ameliorate emphysema in a rat model.

机构信息

Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Mass; Clinical Laboratory Medicine, Faculty of Medical Sciences, Taif University, Taif, Makkah, Saudi Arabia.

Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Mass.

出版信息

J Thorac Cardiovasc Surg. 2023 Jul;166(1):e23-e37. doi: 10.1016/j.jtcvs.2023.03.009. Epub 2023 Mar 17.

DOI:10.1016/j.jtcvs.2023.03.009

PMID:36933786

Abstract

OBJECTIVES

Pulmonary emphysema is characterized by the destruction of alveolar units and reduced gas exchange capacity. In the present study, we aimed to deliver induced pluripotent stem cell-derived endothelial cells and pneumocytes to repair and regenerate distal lung tissue in an elastase-induced emphysema model.

METHODS

We induced emphysema in athymic rats via intratracheal injection of elastase as previously reported. At 21 and 35 days after elastase treatment, we suspended 80 million induced pluripotent stem cell-derived endothelial cells and 20 million induced pluripotent stem cell-derived pneumocytes in hydrogel and injected the mixture intratracheally. On day 49 after elastase treatment, we performed imaging, functional analysis, and collected lungs for histology.

RESULTS

Using immunofluorescence detection of human-specific human leukocyte antigen 1, human-specific CD31, and anti--green fluorescent protein for the reporter labeled pneumocytes, we found that transplanted cells engrafted in 14.69% ± 0.95% of the host alveoli and fully integrated to form vascularized alveoli together with host cells. Transmission electron microscopy confirmed the incorporation of the transplanted human cells and the formation of a blood-air barrier. Human endothelial cells formed perfused vasculature. Computed tomography scans revealed improved vascular density and decelerated emphysema progression in cell-treated lungs. Proliferation of both human and rat cell was higher in cell-treated versus nontreated controls. Cell treatment reduced alveolar enlargement, improved dynamic compliance and residual volume, and improved diffusion capacity.

CONCLUSIONS

Our findings suggest that human induced pluripotent stem cell-derived distal lung cells can engraft in emphysematous lungs and participate in the formation of functional distal lung units to ameliorate the progression of emphysema.

摘要

目的

肺气肿的特征是肺泡单位破坏和气体交换能力降低。在本研究中，我们旨在通过向弹性蛋白酶诱导的肺气肿模型中递送诱导多能干细胞衍生的内皮细胞和肺泡细胞来修复和再生远端肺组织。

方法

我们如先前报道的那样，通过气管内注射弹性酶在无胸腺大鼠中诱导肺气肿。在弹性酶处理后 21 天和 35 天，我们将 8000 万个诱导多能干细胞衍生的内皮细胞和 2000 万个诱导多能干细胞衍生的肺泡细胞悬浮在水凝胶中，并通过气管内注射混合物。在弹性酶处理后第 49 天，我们进行了成像、功能分析，并收集肺组织进行组织学检查。

结果

使用免疫荧光检测人特异性人类白细胞抗原 1、人特异性 CD31 和抗绿色荧光蛋白标记的报告基因，我们发现移植细胞在宿主肺泡的 14.69%±0.95%中定植，并与宿主细胞完全整合形成血管化肺泡。透射电子显微镜证实了移植的人细胞的掺入和血-气屏障的形成。人内皮细胞形成了灌注的血管。计算机断层扫描显示，在细胞治疗的肺中，血管密度增加，肺气肿进展减缓。与未处理对照相比，细胞治疗组的人源和大鼠源细胞增殖更高。细胞治疗减少了肺泡扩大，改善了动态顺应性和残气量，并提高了扩散能力。