Although single-cell RNA sequencing studies have begun to provide compendia of cell expression profiles1,2,3,4,5,6,7,8,9, it has been difficult to systematically identify and localize all molecular cell types in individual organs to create a full molecular cell atlas. Here, using droplet- and plate-based single-cell RNA sequencing of approximately 75,000 human cells across all lung tissue compartments and circulating blood, combined with a multi-pronged cell annotation approach, we create an extensive cell atlas of the human lung. We define the gene expression profiles and anatomical locations of 58 cell populations in the human lung, including 41 out of 45 previously known cell types and 14 previously unknown ones. This comprehensive molecular atlas identifies the biochemical functions of lung cells and the transcription factors and markers for making and monitoring them; defines the cell targets of circulating hormones and predicts local signalling interactions and immune cell homing; and identifies cell types that are directly affected by lung disease genes and respiratory viruses. By comparing human and mouse data, we identified 17 molecular cell types that have been gained or lost during lung evolution and others with substantially altered expression profiles, revealing extensive plasticity of cell types and cell-type-specific gene expression during organ evolution including expression switches between cell types. This atlas provides the molecular foundation for investigating how lung cell identities, functions and interactions are achieved in development and tissue engineering and altered in disease and evolution.
The atlas allowed us to map the cell targets of circulating hormones, based on expression of their cognate receptors. Receptors for some hormones are broadly expressed, indicating direct action throughout the lung (Extended Data Fig. 6a). Other hormones have specific and unexpected targets, such as somatostatin (SSTR1, arteries), melanocortin (MC1R, ionocytes), and oxytocin (OXTR, ciliated cells). Pericytes are predicted targets of several hormones, which could affect their contractile machinery to regulate alveolar perfusion (Extended Data Fig. 6b). Receptors for half the hormones were not detectably expressed so these hormones may not directly influence lung physiology. We also mapped local signalling interactions by examining expression of ligands and receptors, which predicts up to hundreds of interactions among neighbouring cell types (Extended Data Fig. 6c, Supplementary Table 5).
LOLITAS CLUB target
Dot plots showing expression of chemokine receptors (left) and ligands (right) in human lung cells (10x dataset); only cell types and chemokines with detected expression are shown. Colored lines connect ligand sources (target cells) with migrating immune cell types and ionocytes (ion, red) expressing cognate receptor; thicker lines indicate previously unknown interactions. For more details on statistics and reproducibility, see Methods. Bas/Ma, basophil/mast cell; MK, megakaryocyte.
Lung tissue for Tabula Muris Senis40 was obtained as previously described. We obtained additional tissue from two mice expressing Cre recombinase and two expressing oestrogen-inducible Cre recombinase (Cre-ERT2) for conditional cell-specific labelling in vivo with the gene-targeted alleles FVB-Tbx4-LME-cre41,42 (lung stroma) and B6.129-Axin2-cre-ERT241, respectively. Cre-dependent reporter alleles Rosa26ZsGreen1, which expresses cytosolic ZsGreen1 following Cre-mediated recombination, and Rosa26mTmG, which expresses membrane-targeted green fluorescent protein (mGFP) after recombination and membrane-targeted tdTomato (mTomato) in all other tissues, were used to label cells expressing Tbx4 and Axin2, respectively43,44. Induction of the Axin2-cre-ERT2 allele was done by intraperitoneal injection of tamoxifen (3 mg) once a day for three days as described25. All mouse experiments followed applicable regulations and guidelines and were approved by the Institutional Animal Care and Use Committee at Stanford University (Protocol 9780).
We use cookies and other tracking technologies to improve your browsing experience on our site, show personalized content and targeted ads, analyze site traffic, and understand where our audiences come from. To learn more or opt-out, read our Cookie Policy. Please also read our Privacy Notice and Terms of Use, which became effective December 20, 2019.
Methods: In this user centered evaluation, focus groups were conducted with 12 men who had completed the 12-week ViSiT intervention. Participants discussed reasons for participating in and completing the intervention, effects of the intervention, advantages, and areas of improvement of the intervention, and thoughts on the club's involvement. The material was analyzed using thematic analysis according to Braun and Clarke.
Results: The analyses revealed four themes: reasons to participate, motivation and reinforcement, change of habit, and areas for improvement. The intervention was seen as an opportunity to change daily lifestyle behaviors. The group format, as well as the involvement of a prestigious sports club, was important for signing up to the intervention and for motivating continued involvement. The intervention had also resulted in increased knowledge on health and changed mindsets about being more attentive to regulating day-to-day behavior. Although the overall feedback on the intervention was positive, the participants suggested that possibilities to have more individual coaching should be added.
Traditionally, men are underrepresented in community-based weight-loss intervention studies (Pagoto et al., 2012) and in workplace health promotion interventions (Robroek et al., 2009). Also, men appear to be less knowledgeable about risk factors for health and more reluctant than women to engage in health-promoting programmes (Pagoto et al., 2012; Gray et al., 2013; Carroll and Lambe, 2014; Ek, 2015). On this point, there have been several recent initiatives assessing gender-sensitized healthy lifestyle interventions for men performed in contexts such as their work place, with their children or technology-based (Caperchione et al., 2016; Seaton et al., 2017; Williams et al., 2018; Young and Morgan, 2018). Sport venues have been shown to be good locations for gathering people with low health literacy that are not easy to reach with health interventions and health education (Drygas et al., 2013). A new method of attracting men who need to lose weight, a concept called football fans in training (FFIT), has been developed and studied in Scotland. FFIT aims to recruit overweight supporters of professional football clubs and offer them a weight-loss intervention combining evidence-based behavioral change techniques to improve physical activity, sedentary time and diet.
The FFIT intervention uses the traditionally male environment of football clubs, and the powerful social and psychological connections to the team (e.g., belonging, identity, loyalty, validation) that being a supporter creates (Hirt and Clarksson, 2010) along with the opportunity to participate in a men-only group, to maximize men's engagement with an evidence-based, gender-sensitized weight management programme (Wyke, 2012). The FFIT has been designated best practice to affect the lifestyle of the target population by the Canadian Public Health Agency. FFIT has been shown to attract men who were at very high risk of ill health (Hunt et al., 2014a) and shows good results on weight loss (adjusted mean difference between intervention and control 4.9 kg) (Hunt et al., 2014b).
A semi-structured interview guide was created for use in the study. The interview guide contained questions on reasons for participation in the intervention, important effects of the intervention, what made the participants complete the intervention, the participants' perceptions about the success of the intervention, areas for improvement of the intervention and thoughts on the club's involvement in the intervention. Probing questions were used to direct the conversations and get the participants to give detailed answers. Before starting the focus groups the moderator and observer introduced themselves, their role in the research project and interests in the area. In order to make the participants feel relaxed, and to make sure that everyone said something, the first introductory question concerned the best game so far this season for the team they supported. The focus groups were held at the same location as the intervention.
The original FFIT study was carried out in Scotland where football plays an important role in the male culture, and the sport and club context provides an opportunity to reach a large number of men with less favorable health behaviors (Drygas et al., 2013; Hunt et al., 2014a). A similar intervention, Premier League Health, has been carried out in England where the football culture is also strong (Pringle et al., 2013). An adapted version directed to ice hockey fans is carried out in Canada where ice hockey is an important part of being Canadian (Gill et al., 2016). In Sweden, football and hockey are the predominant sports in terms of the number of players and supporters. However, the culture is not as pronounced as in Scotland and Canada. Still, the national top leagues within football and ice hockey are large enough to have an intense supporter culture with loyal members. The results from this study show that the concept of lifestyle interventions promoted by sports clubs seems to attract the target population in Sweden. The participants agreed that both the club involvement and the fact that the intervention was directed to a homogeneous group made them sign up and helped maintain their motivation. From the focus groups, we could not see any differences between the supporters from the football club and ice hockey club. These results are similar to results from FFIT showing that the setting and social situation with likeminded men encouraged the participants and contributed to the desirability to participate in the intervention (Hunt et al., 2014a; Bunn et al., 2016). It has been suggested that associating a health behavior intervention with the commitment and pleasure of being a fan can turn motivation to support the team into motivation to lose weight together with fellow fans (Duncan et al., 2012). 2ff7e9595c
コメント