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Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes

Allergic sensitization to cannabis is an emerging public health concern and is difficult to clinically establish owing to lack of standardized diagnostic approaches. Attempts to develop diagnostic tools were largely hampered by the Schedule I restrictions on cannabis, which limited accessibility for...

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Autores principales: Loblundo, Cali, Severa, Jenna, Peruggia, Gabrielle A., Black, Kristina Reid, Chandra, Suman, Lata, Hemant, ElSohly, Mahmoud, Chapman, Martin D., Deshpande, Deepak A., Nayak, Ajay P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531008/
https://www.ncbi.nlm.nih.gov/pubmed/37762267
http://dx.doi.org/10.3390/ijms241813964
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author Loblundo, Cali
Severa, Jenna
Peruggia, Gabrielle A.
Black, Kristina Reid
Chandra, Suman
Lata, Hemant
ElSohly, Mahmoud
Chapman, Martin D.
Deshpande, Deepak A.
Nayak, Ajay P.
author_facet Loblundo, Cali
Severa, Jenna
Peruggia, Gabrielle A.
Black, Kristina Reid
Chandra, Suman
Lata, Hemant
ElSohly, Mahmoud
Chapman, Martin D.
Deshpande, Deepak A.
Nayak, Ajay P.
author_sort Loblundo, Cali
collection PubMed
description Allergic sensitization to cannabis is an emerging public health concern and is difficult to clinically establish owing to lack of standardized diagnostic approaches. Attempts to develop diagnostic tools were largely hampered by the Schedule I restrictions on cannabis, which limited accessibility for research. Recently, however, hemp was removed from the classified list, and increased accessibility to hemp allows for the evaluation of its practical clinical value for allergy diagnosis. We hypothesized that the proteomic profile is preserved across different cannabis chemotypes and that hemp would be an ideal source of plant material for clinical testing. Using a proteomics-based approach, we examined whether distinct varieties of cannabis plant contain relevant allergens of cannabis. Cannabis extracts were generated from high tetrahydrocannabinol variety (Mx), high cannabidiol variety (V1-19) and mixed profile variety (B5) using a Plant Total Protein Extraction Kit. Hemp extracts were generated using other standardized methods. Protein samples were subjected to nanoscale tandem mass spectrometry. Acquired peptides sequences were examined against the Cannabis sativa database to establish protein identity. Non-specific lipid transfer protein (Can s 3) level was measured using a recently developed ELISA 2.0 assay. Proteomic analysis identified 49 distinct potential allergens in protein extracts from all chemotypes. Most importantly, clinically relevant and validated allergens, such as profilin (Can s 2), Can s 3 and Bet v 1-domain-containing protein 10 (Can s 5), were identified in all chemotypes at label-free quantification (LFP) intensities > 106. However, the oxygen evolving enhancer protein 2 (Can s 4) was not detected in any of the protein samples. Similarly, Can s 2, Can s 3 and Can s 5 peptides were also detected in hemp protein extracts. The validation of these findings using the ELISA 2.0 assay indicated that hemp extract contains 30–37 ng of Can s 3 allergen per µg of total protein. Our proteomic studies indicate that relevant cannabis allergens are consistently expressed across distinct cannabis chemotypes. Further, hemp may serve as an ideal practical substitute for clinical testing, since it expresses most allergens relevant to cannabis sensitization, including the validated major allergen Can s 3.
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spelling pubmed-105310082023-09-28 Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes Loblundo, Cali Severa, Jenna Peruggia, Gabrielle A. Black, Kristina Reid Chandra, Suman Lata, Hemant ElSohly, Mahmoud Chapman, Martin D. Deshpande, Deepak A. Nayak, Ajay P. Int J Mol Sci Article Allergic sensitization to cannabis is an emerging public health concern and is difficult to clinically establish owing to lack of standardized diagnostic approaches. Attempts to develop diagnostic tools were largely hampered by the Schedule I restrictions on cannabis, which limited accessibility for research. Recently, however, hemp was removed from the classified list, and increased accessibility to hemp allows for the evaluation of its practical clinical value for allergy diagnosis. We hypothesized that the proteomic profile is preserved across different cannabis chemotypes and that hemp would be an ideal source of plant material for clinical testing. Using a proteomics-based approach, we examined whether distinct varieties of cannabis plant contain relevant allergens of cannabis. Cannabis extracts were generated from high tetrahydrocannabinol variety (Mx), high cannabidiol variety (V1-19) and mixed profile variety (B5) using a Plant Total Protein Extraction Kit. Hemp extracts were generated using other standardized methods. Protein samples were subjected to nanoscale tandem mass spectrometry. Acquired peptides sequences were examined against the Cannabis sativa database to establish protein identity. Non-specific lipid transfer protein (Can s 3) level was measured using a recently developed ELISA 2.0 assay. Proteomic analysis identified 49 distinct potential allergens in protein extracts from all chemotypes. Most importantly, clinically relevant and validated allergens, such as profilin (Can s 2), Can s 3 and Bet v 1-domain-containing protein 10 (Can s 5), were identified in all chemotypes at label-free quantification (LFP) intensities > 106. However, the oxygen evolving enhancer protein 2 (Can s 4) was not detected in any of the protein samples. Similarly, Can s 2, Can s 3 and Can s 5 peptides were also detected in hemp protein extracts. The validation of these findings using the ELISA 2.0 assay indicated that hemp extract contains 30–37 ng of Can s 3 allergen per µg of total protein. Our proteomic studies indicate that relevant cannabis allergens are consistently expressed across distinct cannabis chemotypes. Further, hemp may serve as an ideal practical substitute for clinical testing, since it expresses most allergens relevant to cannabis sensitization, including the validated major allergen Can s 3. MDPI 2023-09-11 /pmc/articles/PMC10531008/ /pubmed/37762267 http://dx.doi.org/10.3390/ijms241813964 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Loblundo, Cali
Severa, Jenna
Peruggia, Gabrielle A.
Black, Kristina Reid
Chandra, Suman
Lata, Hemant
ElSohly, Mahmoud
Chapman, Martin D.
Deshpande, Deepak A.
Nayak, Ajay P.
Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title_full Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title_fullStr Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title_full_unstemmed Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title_short Proteomics-Based Approach for Detailing the Allergenic Profile of Cannabis Chemotypes
title_sort proteomics-based approach for detailing the allergenic profile of cannabis chemotypes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531008/
https://www.ncbi.nlm.nih.gov/pubmed/37762267
http://dx.doi.org/10.3390/ijms241813964
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