Tsuguhisa Nakayama1, Ivan T Lee2, Wei Le3, Yasuhiro Tsunemi4, Nicole A Borchard3, David Zarabanda3, Sachi S Dholakia3, Philip A Gall3, Angela Yang3, Dayoung Kim3, Makoto Akutsu4, Takashi Kashiwagi4, Zara M Patel3, Peter H Hwang3, Daniel N Frank5, Shin-Ichi Haruna4, Vijay R Ramakrishnan6, Garry P Nolan7, Sizun Jiang8, Jayakar V Nayak9. 1. Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, Calif; Department of Otorhinolaryngology, Jikei University School of Medicine, Tokyo, Japan. 2. Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, Calif; Department of Pathology, Stanford University School of Medicine, Stanford, Calif; Division of Allergy, Immunology, and Rheumatology, Department of Pediatrics, Stanford University School of Medicine, Stanford, Calif. 3. Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, Calif. 4. Department of Otorhinolaryngology-Head and Neck Surgery, Dokkyo Medical University, Tochigi, Japan. 5. Division of Infectious Diseases, University of Colorado, Aurora, Colo. 6. Department of Otolaryngology-Head and Neck Surgery, University of Colorado, Aurora, Colo. 7. Department of Pathology, Stanford University School of Medicine, Stanford, Calif. 8. Department of Pathology, Stanford University School of Medicine, Stanford, Calif; Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Boston, Mass. Electronic address: sjiang3@bidmc.harvard.edu. 9. Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, Calif; Department of Otolaryngology-Head and Neck Surgery, Veterans Affairs Palo Alto Health Care System, Palo Alto, Calif. Electronic address: jnayak@stanford.edu.
Abstract
BACKGROUND: Emerging evidence suggests that chronic rhinosinusitis with nasal polyps (CRSwNP) is a highly heterogeneous disease with disparate inflammatory characteristics between different racial groups and geographies. Currently, little is known about possible underlying distinguishing factors between these inflammatory differences. OBJECTIVE: Our aim was to interrogate differences in CRSwNP disease between White/non-Asian patients and Japanese patients by using whole transcriptome and single-cell RNA gene expression profiling of nasal polyps (NPs). METHODS: We performed whole transcriptome RNA sequencing with endotype stratification of NPs from 8 White patients (residing in the United States) and 9 Japanese patients (residing in Japan). Reproducibility was confirmed by quantitative PCR in an independent validation set of 46 White and 31 Japanese patients. Single-cell RNA sequencing (scRNAseq) was used to stratify key cell types for contributory transcriptional signatures. RESULTS: Unsupervised clustering analysis identified 2 major endotypes that were present within both cohorts of patients with NPs and had previously been reported at the cytokine level: (1) type 2 endotype and (2) non-type 2 endotype. Importantly, there was a statistically significant difference in the proportion of these endotypes between these geographically distinct subgroups with NPs (P = .03). Droplet-based single-cell RNA sequencing further identified prominent type 2 inflammatory transcript expression: C-C motif chemokine ligand 13 (CCL13) and CCL18 in M2 macrophages, as well as cystatin SN (CST1) and CCL26 in basal, suprabasal, and secretory epithelial cells. CONCLUSION: NPs from both racial groups harbor the same 2 major endotypes, which we have determined to be present in differing ratios between each cohort with CRSwNP disease. Distinct inflammatory and epithelial cells contribute to the type 2 inflammatory profiles observed.
BACKGROUND: Emerging evidence suggests that chronic rhinosinusitis with nasal polyps (CRSwNP) is a highly heterogeneous disease with disparate inflammatory characteristics between different racial groups and geographies. Currently, little is known about possible underlying distinguishing factors between these inflammatory differences. OBJECTIVE: Our aim was to interrogate differences in CRSwNP disease between White/non-Asian patients and Japanese patients by using whole transcriptome and single-cell RNA gene expression profiling of nasal polyps (NPs). METHODS: We performed whole transcriptome RNA sequencing with endotype stratification of NPs from 8 White patients (residing in the United States) and 9 Japanese patients (residing in Japan). Reproducibility was confirmed by quantitative PCR in an independent validation set of 46 White and 31 Japanese patients. Single-cell RNA sequencing (scRNAseq) was used to stratify key cell types for contributory transcriptional signatures. RESULTS: Unsupervised clustering analysis identified 2 major endotypes that were present within both cohorts of patients with NPs and had previously been reported at the cytokine level: (1) type 2 endotype and (2) non-type 2 endotype. Importantly, there was a statistically significant difference in the proportion of these endotypes between these geographically distinct subgroups with NPs (P = .03). Droplet-based single-cell RNA sequencing further identified prominent type 2 inflammatory transcript expression: C-C motif chemokine ligand 13 (CCL13) and CCL18 in M2 macrophages, as well as cystatin SN (CST1) and CCL26 in basal, suprabasal, and secretory epithelial cells. CONCLUSION: NPs from both racial groups harbor the same 2 major endotypes, which we have determined to be present in differing ratios between each cohort with CRSwNP disease. Distinct inflammatory and epithelial cells contribute to the type 2 inflammatory profiles observed.