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Assessment of Changes in Genetic Transcriptome in Nasal Epithelial Cells Exposed to Black Carbon and Pollen Allergen by High-throughput Transcriptomics
Luo Zhang
Beijing Tongren Hospital
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0910-9884
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Evidence suggests that air pollution may be associated with an increase in airway responsiveness to allergens, an increase in bioavailability of airborne allergens and possibly exacerbation of allergic rhinitis (AR). Environmental black carbon (BC) is an important constituent of atmospheric particulate matter (PM), for which the mechanisms underlying its effects have not been fully elucidated in AR. The objective of the present study was to determine the BC and pollen-induced alterations in the transcriptome in human nasal epithelial cells (hNECs) in vitro.
Methods: hNECs were prepared from nasal epithelial mucosal samples of healthy individuals undergoing nasal surgery (turbinoplasty or septoplasty). The hNECs were established as air-liquid interface (ALI) cultures and exposed to BC alone or in combination with pollen allergen. The changes in the transcriptome were analyzed by high-throughput RNA sequencing (RNA-Seq). Some of the differentially expressed genes were verified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Enrichment analysis based on Gene Ontology (GO) and KEGG database for each Gene was performed to determine their major biological functions and pathways.
Results: Exposure to ≥50 μg/ml BC or 25μg/ml BC + 200g /ml pollen significantly decreased cell viability in the hNECs compared to control (p<0.05) or 25μg/ml BC alone (p<0.05). Expression of 114 genes (including 80 up-regulated and 34 down-regulated) and 293 genes (including 233 up-regulated and 60 down-regulated genes) was significantly altered following exposure to 200 μg/ml BC and 25 μg/ml BC =200 μg/ml pollen respectively. For 200 μg/ml BC, up-regulated GO terms were mainly associated with hypoxia stress response, whereas for 25 μg/ml BC + 200 μg/ml pollen treatment the top enriched GO terms were associated with inflammatory response including regulation of neutrophil migration and chemotaxis, macrophage differentiation and chemotaxis, mast cell activation, and phagocytosis. KEGG pathway analysis indicated the top 10 upstream regulators to be IL1B, CSF1, CCL2, TLR2, LPL, IGF8, SPP1, CXCL8, FCER1G and IL1RN. The expressions of inflammation related gene IL1B, CSF1 and FCER1G were elevated as measured by RT-qPCR assay.
Conclusion: BC and pollen allergen may induce innate immune and allergic inflammation in hNECs, and therefore potentially exacerbate the symptoms of AR in affected individuals.
Keywords
Black carbon, Pollen allergen, Differential gene expression, Gene Ontology, High-throughput RNA sequencing, Human nasal epithelial cell cultures (hNECs), inflammation, KEGG pathway analysis, Transcriptome
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CURRENT STATUS: Under Review
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Posted 04 Dec, 2020
No community comments so far
Review #2 received
Received 23 Jan, 2021
Review #1 received
Received 23 Jan, 2021
Reviewer #2 agreed
On 14 Jan, 2021
Reviewer #1 agreed
On 13 Jan, 2021
Reviewers invited
Invitations sent on 20 Dec, 2020
Editor assigned
On 27 Nov, 2020
Submission checks complete
On 27 Nov, 2020
Editor invited
On 27 Nov, 2020
First submitted
On 26 Nov, 2020
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Subject Areas
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This preprint is under consideration at Allergy, Asthma & Clinical Immunology. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Assessment of Changes in Genetic Transcriptome in Nasal Epithelial Cells Exposed to Black Carbon and Pollen Allergen by High-throughput Transcriptomics
Luo Zhang
Beijing Tongren Hospital
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0910-9884
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 04 Dec, 2020
No community comments so far
Review #2 received
Received 23 Jan, 2021
Review #1 received
Received 23 Jan, 2021
Reviewer #2 agreed
On 14 Jan, 2021
Reviewer #1 agreed
On 13 Jan, 2021
Reviewers invited
Invitations sent on 20 Dec, 2020
Editor assigned
On 27 Nov, 2020
Submission checks complete
On 27 Nov, 2020
Editor invited
On 27 Nov, 2020
First submitted
On 26 Nov, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Allergy & Immune Disorders
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Evidence suggests that air pollution may be associated with an increase in airway responsiveness to allergens, an increase in bioavailability of airborne allergens and possibly exacerbation of allergic rhinitis (AR). Environmental black carbon (BC) is an important constituent of atmospheric particulate matter (PM), for which the mechanisms underlying its effects have not been fully elucidated in AR. The objective of the present study was to determine the BC and pollen-induced alterations in the transcriptome in human nasal epithelial cells (hNECs) in vitro.
Methods: hNECs were prepared from nasal epithelial mucosal samples of healthy individuals undergoing nasal surgery (turbinoplasty or septoplasty). The hNECs were established as air-liquid interface (ALI) cultures and exposed to BC alone or in combination with pollen allergen. The changes in the transcriptome were analyzed by high-throughput RNA sequencing (RNA-Seq). Some of the differentially expressed genes were verified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Enrichment analysis based on Gene Ontology (GO) and KEGG database for each Gene was performed to determine their major biological functions and pathways.
Results: Exposure to ≥50 μg/ml BC or 25μg/ml BC + 200g /ml pollen significantly decreased cell viability in the hNECs compared to control (p<0.05) or 25μg/ml BC alone (p<0.05). Expression of 114 genes (including 80 up-regulated and 34 down-regulated) and 293 genes (including 233 up-regulated and 60 down-regulated genes) was significantly altered following exposure to 200 μg/ml BC and 25 μg/ml BC =200 μg/ml pollen respectively. For 200 μg/ml BC, up-regulated GO terms were mainly associated with hypoxia stress response, whereas for 25 μg/ml BC + 200 μg/ml pollen treatment the top enriched GO terms were associated with inflammatory response including regulation of neutrophil migration and chemotaxis, macrophage differentiation and chemotaxis, mast cell activation, and phagocytosis. KEGG pathway analysis indicated the top 10 upstream regulators to be IL1B, CSF1, CCL2, TLR2, LPL, IGF8, SPP1, CXCL8, FCER1G and IL1RN. The expressions of inflammation related gene IL1B, CSF1 and FCER1G were elevated as measured by RT-qPCR assay.
Conclusion: BC and pollen allergen may induce innate immune and allergic inflammation in hNECs, and therefore potentially exacerbate the symptoms of AR in affected individuals.
Figure 1
Figure 1
Figure 2
Figure 2
Figure 3
Figure 3
Figure 4
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