Platform Effects on Regeneration by Pulmonary Basal Cells as Evaluated by Single-Cell RNA Sequencing

Greaney, Allison M.; Adams, Taylor; Raredon, Micha Sam Brickman; Gubbins, Elise; Schupp, Jonas C.; Engler, Alexander J.; Ghaedi, Mahboobe; Yuan, Yifan; Kaminski, Naftali; Niklason, Laura E.

doi:10.1016/j.celrep.2020.03.004

Cited by 38 publications

(33 citation statements)

References 100 publications

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“…Recellularization of a decellularized lung with endoderm cells appears to replicate airway epithelial wound repair and regeneration instead of lung development. Our findings support the use of pulmonary basal cells as a cell source for reseeding an acellular lung 45 , 69 . The recent finding that acellular lung scaffolds support region-specific epithelial differentiation of primary TP63 + basal cells better than artificial platforms like organoids or air-liquid-interface cultures 45 emphasizes this concept.…”

Section: Discussionsupporting

confidence: 82%

“…Multipotent stem cells are an ideal cell source for reseeding acellular scaffolds, specifically those of complex multi-cellular organs like the lung. Various studies have shown that pre-differentiated lung progenitors under specific supplementation conditions can adhere, proliferate and differentiate to airway epithelium on acellular lung scaffolds 2 , 44 , 45 . We have shown that acellular lung ECM without supplementation promotes endoderm differentiation into proximal airway cells 8 .…”

Section: Discussionmentioning

confidence: 99%

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TP63 basal cells are indispensable during endoderm differentiation into proximal airway cells on acellular lung scaffolds

et al. 2021

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The use of decellularized whole-organ scaffolds for bioengineering of organs is a promising avenue to circumvent the shortage of donor organs for transplantation. However, recellularization of acellular scaffolds from multicellular organs like the lung with a variety of different cell types remains a challenge. Multipotent cells could be an ideal cell source for recellularization. Here we investigated the hierarchical differentiation process of multipotent ES-derived endoderm cells into proximal airway epithelial cells on acellular lung scaffolds. The first cells to emerge on the scaffolds were TP63+ cells, followed by TP63+/KRT5+ basal cells, and finally multi-ciliated and secretory airway epithelial cells. TP63+/KRT5+ basal cells on the scaffolds simultaneously expressed KRT14, like basal cells involved in airway repair after injury. Removal of TP63 by CRISPR/Cas9 in the ES cells halted basal and airway cell differentiation on the scaffolds. These findings suggest that differentiation of ES-derived endoderm cells into airway cells on decellularized lung scaffolds proceeds via TP63+ basal cell progenitors and tracks a regenerative repair pathway. Understanding the process of differentiation is key for choosing the cell source for repopulation of a decellularized organ scaffold. Our data support the use of airway basal cells for repopulating the airway side of an acellular lung scaffold.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

TP63 basal cells are indispensable during endoderm differentiation into proximal airway cells on acellular lung scaffolds

et al. 2021

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“…Investigation of stem cell activity in adult lung homeostasis and regeneration is a rapidly expanding field with solid evidence for cellular regeneration in adult humans 29 . A number of studies have explored the stem cell function of alveolar type 2 and basal cells expressing KRT5 in different contexts, mostly relying on in-vitro or animal models 30 – 33 ; while others have used scRNAseq technologies to gain insights on basal cell differentiation dynamics in different in-vitro settings 34 and on neuroendocrine cell regeneration in the adult mouse lung 35 . Collectively, these studies have generated extensive insight into the alveolar epithelium lineage; however, a systems-wise, comprehensively defined lineage is yet to be identified.…”

Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 receptor is co-expressed with elements of the kinin–kallikrein, renin–angiotensin and coagulation systems in alveolar cells

Sidarta-Oliveira

Jara

Ferruzzi

et al. 2020

Sci Rep

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SARS-CoV-2, the pathogenic agent of COVID-19, employs angiotensin converting enzyme-2 (ACE2) as its cell entry receptor. Clinical data reveal that in severe COVID-19, SARS-CoV-2 infects the lung, leading to a frequently lethal triad of respiratory insufficiency, acute cardiovascular failure, and coagulopathy. Physiologically, ACE2 plays a role in the regulation of three systems that could potentially be involved in the pathogenesis of severe COVID-19: the kinin–kallikrein system, resulting in acute lung inflammatory edema; the renin–angiotensin system, promoting cardiovascular instability; and the coagulation system, leading to thromboembolism. Here we assembled a healthy human lung cell atlas meta-analysis with ~ 130,000 public single-cell transcriptomes and show that key elements of the bradykinin, angiotensin and coagulation systems are co-expressed with ACE2 in alveolar cells and associated with their differentiation dynamics, which could explain how changes in ACE2 promoted by SARS-CoV-2 cell entry result in the development of the three most severe clinical components of COVID-19.

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“…In addition, we observed a sustained increase in the pro‐regenerative airway phenotypic marker KRT14 in all patients after lifting to ALI and consistent decreases in all patients for the basal cell marker KRT5 on rECM. [ 15 ] Finally, we tested whether primary HBECs survive 3D FRESH bioprinting, as primary cells are known to be more sensitive than cell lines. In line with our previous data (Figure 1m), we observed a significantly higher number of viable cells present in the rECM hydrogel (≈90%) as compared to alginate (≈60%) after bioprinting (Figure 3e) with homogenous distribution and viability up to 35 days (Figure 3f).…”

Section: Figurementioning

confidence: 99%

Extracellular‐Matrix‐Reinforced Bioinks for 3D Bioprinting Human Tissue

et al. 2020

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Recent advances in 3D bioprinting allow for generating intricate structures with dimensions relevant for human tissue, but suitable bioinks for producing translationally relevant tissue with complex geometries remain unidentified. Here, a tissue‐specific hybrid bioink is described, composed of a natural polymer, alginate, reinforced with extracellular matrix derived from decellularized tissue (rECM). rECM has rheological and gelation properties beneficial for 3D bioprinting while retaining biologically inductive properties supporting tissue maturation ex vivo and in vivo. These bioinks are shear thinning, resist cell sedimentation, improve viability of multiple cell types, and enhance mechanical stability in hydrogels derived from them. 3D printed constructs generated from rECM bioinks suppress the foreign body response, are pro‐angiogenic and support recipient‐derived de novo blood vessel formation across the entire graft thickness in a murine model of transplant immunosuppression. Their proof‐of‐principle for generating human tissue is demonstrated by 3D bioprinting human airways composed of regionally specified primary human airway epithelial progenitor and smooth muscle cells. Airway lumens remained patent with viable cells for one month in vitro with evidence of differentiation into mature epithelial cell types found in native human airways. rECM bioinks are a promising new approach for generating functional human tissue using 3D bioprinting.

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Platform Effects on Regeneration by Pulmonary Basal Cells as Evaluated by Single-Cell RNA Sequencing

Cited by 38 publications

References 100 publications

TP63 basal cells are indispensable during endoderm differentiation into proximal airway cells on acellular lung scaffolds

TP63 basal cells are indispensable during endoderm differentiation into proximal airway cells on acellular lung scaffolds

SARS-CoV-2 receptor is co-expressed with elements of the kinin–kallikrein, renin–angiotensin and coagulation systems in alveolar cells

Extracellular‐Matrix‐Reinforced Bioinks for 3D Bioprinting Human Tissue

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