Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis
Authors: Kenichi Okuda
Burton F Dickey
Christopher P Crum
Wanda K O&#x27;Neal
Kalpaj R Parekh
Marcin Duleba
Matthew Vincent
John F Engelhardt
Soma S K Jyothula
Suchan Niroula
Shan Wang
Wei Wang
Michael E Wechsler
Rahul Neupane
Wa Xian
Rajasekaran Mahalingam
Harry Karmouty-Quintana
Mark L Metersky
Audrey-Ann Liew
Frank D McKeon
Omar Ibrahim
Richard C Boucher
Jingzhong Xie
Kristina Goller
Wei Rao
Tinne C J Mertens
Journal: Cell (2020)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: COPD; chronic lung disease; fibrosis; inflammation; lung
Affiliations: Stem Cell Center, Department of Biology and Biochemistry, University of Houston, Houston, TX 77003, USA., Nüwa Medical Systems, Houston, TX 77479, USA., Marsico Lung Center, University of North Carolina, Chapel Hill, NC 27599, USA., Department of Pulmonary Medicine, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA., Department of Medicine, National Jewish Health, Denver, CO 80206, USA., Department of Medicine, Division of Pulmonary, Critical Care, and Sleep Medicine, University of Connecticut School of Medicine, Farmington, CT 06032, USA., Department of Anatomy and Cell Biology, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA., Department of Biochemistry and Molecular Biology, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030, USA., Department of Internal Medicine, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030, USA., Department of Pathology, Harvard Medical School and Brigham and Women's Hospital, Boston, MA 02215, USA., Department of Anatomy and Cell Biology, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA; Department of Surgery, Division of Cardiothoracic Surgery, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA., Stem Cell Center, Department of Biology and Biochemistry, University of Houston, Houston, TX 77003, USA. Electronic address: .

Abstract:  Chronic obstructive pulmonary disease (COPD) is a progressive condition of chronic bronchitis, small airway obstruction, and emphysema that represents a leading cause of death worldwide. While inflammation, fibrosis, mucus hypersecretion, and metaplastic epithelial lesions are hallmarks of this disease, their origins and dependent relationships remain unclear. Here we apply single-cell cloning technologies to lung tissue of patients with and without COPD. Unlike control lungs, which were dominated by normal distal airway progenitor cells, COPD lungs were inundated by three variant progenitors epigenetically committed to distinct metaplastic lesions. When transplanted to immunodeficient mice, these variant clones induced pathology akin to the mucous and squamous metaplasia, neutrophilic inflammation, and fibrosis seen in COPD. Remarkably, similar variants pre-exist as minor constituents of control and fetal lung and conceivably act in normal processes of immune surveillance. However, these same variants likely catalyze the pathologic and progressive features of COPD when expanded to high numbers. 

Retracted: No

PubPeer comments:
 **The localization of IL-33 and IL-1 beta in Figure 5 D looks very similar.** **What antibodies did authors use for immunofluorescence micrographs?** ![file] , 
 There is no information regarding anti-IL33 and IL-1 beta antibodies. Could the authors please clarify? ![file] , 
 #2 Thanks. We will , 
 ![file] , 
 #4 Thanks for your question. This staining was done using both Muc5Ac and Muc5B antibodies on the same sections showing negative stainings on the CLST_1 differentiated epithelia, that s why they re similar, actually, the same. , 
 #1 Thanks for pointing that out. We unintentionally put the same images as IL1b for IL-33 by mistake during the revision processes. We already contacted the editor for the correction. See the correct IL-33 staining image.![file] , 
 #6 It looks like you did not put the same images as IL1b for IL-33 by mistake. Please check the yellow circle in the attached image. The two pictures are not entirely the same. Furthermore, you added the horizontal rotation to the image. How would you describe them? ![file] , 
 CLST3 (yellow box) and CLST4 (green box) are different samples, but they have similar nuclear positions. ![file] , 
 #8 Interestingly, CLST3 and CLST4 are forming similar structures in vitro upon air-liquid interface differentiation, both squamous cell metaplasia. They may look alike, though are different sections from respective ALI structures, see the original images before croppings: ![file] , 
 #4 Thank you for your reply. *Thanks for your question. This staining was done using both Muc5Ac and Muc5B antibodies on the same sections showing negative stainings on the CLST_1 differentiated epithelia, that s why they re similar, actually, the same.* But, as staining antibodies of MUC5AC and MUC5B are same host (rabbit), if stained, you could not distinguish which antibodies by using Alexa594 labelled anti-rabbit antibody on same section. Therefore, it considered that this protocol is not adequate. Also, horizontal rotation of Figure S1C upper and lower seems a bit artificial. So description of Figure S1C is not in this article, It is thought that it s better to delete these sentences. ![file] , 
 **Comment (1)**: I was also concerned that pictures of IL1b and IL-33 would not match exactly, although you claimed they were the same. Could you please tell us your opinion about the comment from #7? **Comment (2)**: I understand that CLST3 and CLST4 are forming similar structures with squamous cell metaplasia-like phenotype in ALI experiments. However, as you deeply understand, there is no same cell position in culture experiments, even if the conditions are identical. Please check not only the nuclear position but also the background in the attached image. ***Your additional original images before cropping support #8u2019s comment more strongly. The CLST3 (magenta box: SCGB1A1 SFTPB) and CLST4 (orange box: IVL) seem to be serial sections in your results***. ![file] ,