Roya Babaei-Jadidi1, Arundhati Dongre1, Suzanne Miller2, Marcos Castellanos Uribe3, Iain D Stewart4, Zoe M Thompson1, Abdolrahman S Nateri5, Peter Bradding6, Sean T May7, Debbie Clements8, Simon R Johnson9. 1. University of Nottingham, Division of Respiratory Medicine, Nottingham, United Kingdom of Great Britain and Northern Ireland. 2. University of Nottingham, 6123, Division of Respiratory Medicine and Respiratory Research Unit, Nottingham, United Kingdom of Great Britain and Northern Ireland. 3. University of Nottingham, 6123, Nottingham Arabidopsis Stock Centre, , Nottingham, United Kingdom of Great Britain and Northern Ireland. 4. University of Nottingham Faculty of Medicine and Health Sciences, 12207, Nottingham, United Kingdom of Great Britain and Northern Ireland. 5. University of Nottingham, 6123, Cancer Genetics & Stem Cell Group, Biodiscovery Institute, Nottingham, United Kingdom of Great Britain and Northern Ireland. 6. Leicester Institute for Lung Health, Department of Infection, Immunity and Inflammation, Leicester, United Kingdom of Great Britain and Northern Ireland. 7. University of Nottingham, 6123, Nottingham Arabidopsis Stock Centre, Nottingham, United Kingdom of Great Britain and Northern Ireland. 8. University of Nottingham, Division of Respiratory Medicine and Biodiscovery Institute, Nottingham, United Kingdom of Great Britain and Northern Ireland. 9. University of Nottingham, Division of Respiratory Medicine, Biodiscovery Institute and NIHR Biomedical Research Centre, Nottingham, United Kingdom of Great Britain and Northern Ireland; simon.johnson@nottingham.ac.uk.
Abstract
RATIONALE: Lymphangioleiomyomatosis is a multisystem disease causing lung cysts and respiratory failure. Loss of tuberous sclerosis complex (TSC) gene function results in a clone of 'LAM cells' with dysregulated mTOR activity. LAM cells and fibroblasts form lung nodules that also contain mast cells although their significance is unknown. OBJECTIVES: To understand the mechanism of mast cell accumulation and their role in the pathogenesis of LAM. Methods, Measurements and Main Results: Transcriptional profiling, quantitative RT-PCR and ELISA showed that LAM derived cell / fibroblast co-cultures induced multiple CXC chemokines in fibroblasts. Compared with normal tissue, LAM lungs had increased tryptase positive mast cells expressing CXC chemokine receptors (p<0.05). Mast cells located around the periphery of LAM nodules were positively associated with rate of lung function loss (p=0.016). In vitro, LAM spheroid TSC2 null cell / fibroblast co-cultures attracted mast cells, which was inhibited by pharmacologic and CRISPR-cas9 inhibition of CXCR1 and 2. LAM spheroids caused mast cell tryptase release, which induced fibroblast proliferation and increased LAM spheroid size (1.36±0.24 fold, p=0.0019). The tryptase inhibitor APC366 and sodium cromoglycate inhibited mast cell induced spheroid growth. Using an immuno-competent Tsc2 null murine homograft model, sodium cromoglycate markedly reduced mast cell activation and Tsc2 null lung tumour burden (vehicle: 32.5.3%±23.6 and cromoglycate: 5.5%±4.3. p=0.0035). CONCLUSIONS: LAM cell / fibroblast interactions attract mast cells where tryptase release contributes to disease progression. Repurposing sodium cromoglycate for use in LAM should be studied as an alternative or adjunct to mTOR inhibitor therapy.
RATIONALE: Lymphangioleiomyomatosis is a multisystem disease causing lung cysts and respiratory failure. Loss of tuberous sclerosis complex (TSC) gene function results in a clone of 'LAM cells' with dysregulated mTOR activity. LAM cells and fibroblasts form lung nodules that also contain mast cells although their significance is unknown. OBJECTIVES: To understand the mechanism of mast cell accumulation and their role in the pathogenesis of LAM. Methods, Measurements and Main Results: Transcriptional profiling, quantitative RT-PCR and ELISA showed that LAM derived cell / fibroblast co-cultures induced multiple CXC chemokines in fibroblasts. Compared with normal tissue, LAM lungs had increased tryptase positive mast cells expressing CXC chemokine receptors (p<0.05). Mast cells located around the periphery of LAM nodules were positively associated with rate of lung function loss (p=0.016). In vitro, LAM spheroid TSC2 null cell / fibroblast co-cultures attracted mast cells, which was inhibited by pharmacologic and CRISPR-cas9 inhibition of CXCR1 and 2. LAM spheroids caused mast cell tryptase release, which induced fibroblast proliferation and increased LAM spheroid size (1.36±0.24 fold, p=0.0019). The tryptase inhibitor APC366 and sodium cromoglycate inhibited mast cell induced spheroid growth. Using an immuno-competent Tsc2 null murine homograft model, sodium cromoglycate markedly reduced mast cell activation and Tsc2 null lung tumour burden (vehicle: 32.5.3%±23.6 and cromoglycate: 5.5%±4.3. p=0.0035). CONCLUSIONS:LAM cell / fibroblast interactions attract mast cells where tryptase release contributes to disease progression. Repurposing sodium cromoglycate for use in LAM should be studied as an alternative or adjunct to mTOR inhibitor therapy.
Authors: Jesse D Kirkpatrick; Ava P Soleimany; Jaideep S Dudani; Heng-Jia Liu; Hilaire C Lam; Carmen Priolo; Elizabeth P Henske; Sangeeta N Bhatia Journal: Eur Respir J Date: 2022-04-14 Impact factor: 33.795
Authors: Carmen Herranz; Francesca Mateo; Alexandra Baiges; Gorka Ruiz de Garibay; Alexandra Junza; Simon R Johnson; Suzanne Miller; Nadia García; Jordi Capellades; Antonio Gómez; August Vidal; Luis Palomero; Roderic Espín; Ana I Extremera; Eline Blommaert; Eva Revilla-López; Berta Saez; Susana Gómez-Ollés; Julio Ancochea; Claudia Valenzuela; Tamara Alonso; Piedad Ussetti; Rosalía Laporta; Antoni Xaubet; José A Rodríguez-Portal; Ana Montes-Worboys; Carlos Machahua; Jaume Bordas; Javier A Menendez; Josep M Cruzado; Roser Guiteras; Christophe Bontoux; Concettina La Motta; Aleix Noguera-Castells; Mario Mancino; Enrique Lastra; Raúl Rigo-Bonnin; Jose C Perales; Francesc Viñals; Alvaro Lahiguera; Xiaohu Zhang; Daniel Cuadras; Coline H M van Moorsel; Joanne J van der Vis; Marian J R Quanjel; Harilaos Filippakis; Razq Hakem; Chiara Gorrini; Marc Ferrer; Aslihan Ugun-Klusek; Ellen Billett; Elżbieta Radzikowska; Álvaro Casanova; María Molina-Molina; Antonio Roman; Oscar Yanes; Miquel A Pujana Journal: EMBO Mol Med Date: 2021-08-11 Impact factor: 12.137