Gas exchange in the lung occurs within alveoli, air-filled sacs composed of type 2 and type 1 epithelial cells (AEC2s and AEC1s), capillaries, and various resident mesenchymal cells. Here, we use a combination of in vivo clonal lineage analysis, different injury/repair systems, and in vitro culture of purified cell populations to obtain new information about the contribution of AEC2s to alveolar maintenance and repair. Genetic lineage-tracing experiments showed that surfactant protein C-positive (SFTPC-positive) AEC2s self renew and differentiate over about a year, consistent with the population containing long-term alveolar stem cells. Moreover, if many AEC2s were specifically ablated, high-resolution imaging of intact lungs showed that individual survivors undergo rapid clonal expansion and daughter cell dispersal. Individual lineage-labeled AEC2s placed into 3D culture gave rise to self-renewing "alveolospheres," which contained both AEC2s and cells expressing multiple AEC1 markers, including HOPX, a new marker for AEC1s. Growth and differentiation of the alveolospheres occurred most readily when cocultured with primary PDGFRα⁺ lung stromal cells. This population included lipofibroblasts that normally reside close to AEC2s and may therefore contribute to a stem cell niche in the murine lung. Results suggest that a similar dynamic exists between AEC2s and mesenchymal cells in the human lung.

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Type 2 alveolar cells are stem cells in adult lung

Background. The gas nitric oxide (NO) is an important endothelium-derived relaxing factor, inactivated by rapid combination with heme in hemoglobin. Methods and Results. Awake spontaneously breathing lambs inhaled 5-80 ppm NO with an acutely constricted pulmonary circulation due to either infusion of the stable thromboxane endoperoxide analogue U46619 or breathing a hypoxic gas mixture. Within 3 minutes after adding 40 ppm NO or more to inspired gas, pulmonary hypertension was reversed. Systemic vasodilation did not occur. Pulmonary hypertension resumed within 3-6 minutes of ceasing NO inhalation. During U46619 infusion pulmonary vasodilation was maintained up to 1 hour without tolerance. In the normal lamb, NO inhalation produced no hemodynamic changes. Breathing 80 ppm NO for 3 hours did not increase either methemoglobin or extravascular lung water levels nor modify lung histology compared with control lambs. Conclusions. Low dose inhaled NO (5-80 ppm) is a selective pulmonary vasodilator reversing both hypoxia- and thromboxane-induced pulmonary hypertension in the awake lamb [corrected].

Inhaled nitric oxide. A selective pulmonary vasodilator reversing hypoxic pulmonary vasoconstriction.

Alveoli are gas-exchange sacs lined by squamous alveolar type (AT) 1 cells and cuboidal, surfactant-secreting AT2 cells. Classical studies suggested that AT1 arise from AT2 cells, but recent studies propose other sources. Here we use molecular markers, lineage tracing and clonal analysis to map alveolar progenitors throughout the mouse lifespan. We show that, during development, AT1 and AT2 cells arise directly from a bipotent progenitor, whereas after birth new AT1 cells derive from rare, self-renewing, long-lived, mature AT2 cells that produce slowly expanding clonal foci of alveolar renewal. This stem-cell function is broadly activated by AT1 injury, and AT2 self-renewal is selectively induced by EGFR (epidermal growth factor receptor) ligands in vitro and oncogenic Kras(G12D) in vivo, efficiently generating multifocal, clonal adenomas. Thus, there is a switch after birth, when AT2 cells function as stem cells that contribute to alveolar renewal, repair and cancer. We propose that local signals regulate AT2 stem-cell activity: a signal transduced by EGFR-KRAS controls self-renewal and is hijacked during oncogenesis, whereas another signal controls reprogramming to AT1 fate.

/pdf/alveolar-progenitor-and-stem-cells-in-lung-development-1cg2vfe08h.pdf

Alveolar progenitor and stem cells in lung development, renewal and cancer

The Role of Scgb1a1+ Clara Cells in the Long-Term Maintenance and Repair of Lung Airway, but Not Alveolar, Epithelium

Repair and regeneration of the respiratory system: complexity, plasticity, and mechanisms of lung stem cell function.

Transformation of alveolar type 2 cells to type 1 cells following exposure to NO2.

This research was undertaken to study the kinetics of Type 2 cell division and the fate of the Type 2 sister cell following exposure to NO(2). To accomplish this, male rats were exposed to NO(2). Dividing cells were labeled with (3)H-TdR and studied with autoradiographic technics in the light and electron microscopes for up to 72 hours after labeling. The kinetics of cell division were determined from a curve constructed from the percent of labeled mitotic figures. The fate of the Type 2 sister cells was determined by studying tissues at 24, 48 and 72 hours after labeling with (3)H-TdR. The results show that Type 2 cells may divide and the sister cells transform into Type 1 cells. These data support an interpretation of the mechanism for cell renewal of the alveolar epithelium in which Type 2 cells are the progenitor cells for Type 1 cells.

Renewal of alveolar epithelium in the rat following exposure to NO2.

Significant changes in the lungs of rats exposed to nitrogen dioxide occur during the first 72 hours in animals exposed to 17 ppm. These changes appear in focal areas at the level of the terminal bronchiole: loss of cilia; thickening of tissue; injury to the epithelium lining the adveoli adjacent to the terminal bronchioles, when whole type 1 cells slough away leaving the basement membrane exposed to the air; between 24 and 48 hours of continuous exposure, these areas are repaired with a low cuboidal cell type that tolerates NO2 and thickens the air-blood barrier; and eventual formation of crystalloids and interruption of ciliogenesis. Changes at 2 ppm include loss of cilia, hypertrophy and focal hyperplasia in the epithelium of the terminal bronchiole, and “apparent” return to normal after 21 days of continuous exposure.

Early response of lungs to low levels of nitrogen dioxide. Light and electron microscopy.

The epithelial response in the terminal bronchioles and alveoli was examined after exposure to approximately 05 and 09 ppm ozone (O3) Loss of ciliated cells from the terminal bronchiole was pronounced after 6 to 10 hours of exposure; however, nonciliated cells were resistant to injury from O3 In contrast, type 1 alveolar lining cells were very sensitive to O2 and were severely damaged or removed from significant areas of the proximal alveoli after as little as 2 hours of exposure to 05 ppm The basement lamina was left devoid of an epithelium with the exception of type 2 cells, which were very resistant and retained a relatively normal appearance Under continuous exposure, repair of the lesion area was accomplished in approximately 48 hours through proliferation of the type 2 cells The new cells were cuboidal at first but subsequently became more squamous in appearance Exposure for 8 to 10 hours followed by residence in clean air resulted in a proliferative response at 48 hours, about equal to that observed after continuous exposure

Early response of lung to low levels of ozone

Effects of continuous exposure to low concentrations of NO2 on cell proliferation in bronchiolar and alveolar tissues were determined. Young rats were exposed continuously for as long as 360 days, removed at intervals, and killed. Dividing cells were labeled with tritiated thymidine. Animals exposed to 17 ppm NO2 showed an Increased labeling index in terminal bronchioles and alveoli after two to three days of exposure; the index returned to control levels by five days. Similar results were found in animals exposed to 2 ppm NO2, but the increase was not as great. This resulted in epithelial hyperplasia of terminal bronchioles and increased cellularity of alveoli associated with the distal ends of the terminal bronchioles. In peripheral alveoli, there was an increase in turnover rate of type 2 alveolar cells. All of these changes occurred within three days of continuous exposure. After this time, the areas of cellularity did not increase In size and the labeling indexes returned to normal for the remainder ...

Gustave Freeman

Papers

Transformation of alveolar type 2 cells to type 1 cells following exposure to NO2.

Renewal of alveolar epithelium in the rat following exposure to NO2.

Early response of lungs to low levels of nitrogen dioxide. Light and electron microscopy.

Early response of lung to low levels of ozone

Cell renewal in the lungs of rats exposed to low levels of NO2.