360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens
OBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Cambridge University Press
2023-04-01
|
| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866123004004/type/journal_article |
| _version_ | 1797840485596790784 |
|---|---|
| author | Wilfrido Mojica Bei Yang Chang Chieh Hsu Yun Wu Alexandra Izydorczak Troy D. Wood Dara Cho Natesh Parashurama Donald Yergeau Supriya D Mahajan |
| author_facet | Wilfrido Mojica Bei Yang Chang Chieh Hsu Yun Wu Alexandra Izydorczak Troy D. Wood Dara Cho Natesh Parashurama Donald Yergeau Supriya D Mahajan |
| author_sort | Wilfrido Mojica |
| collection | DOAJ |
| description | OBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can contribute to the further development of Precision Medicine. METHODS/STUDY POPULATION: Transurethral resection of bladder tumors were selected for testing based on availability and tissue composition. A wash step was used to generate daughter aliquots composed of dislodged cells and a solution with prior contact to the parent tissue. This wash step served two purposes: 1) reduce the amount of contaminating material from spreading to other cases, a problem known to be associated with this type of specimen; and 2) create aliquots from which additional informative data could be generated. These daughter aliquots were then examined to determine their value as a source for exosome profiling, metabolomic studies, molecular characterization and organoid development. The parent tissue was not compromised, was able to undergo conventional processing and yielded results equivalent to unwashed specimens. RESULTS/ANTICIPATED RESULTS: Exosomes secreted by the tumor cells were identified to be present in the daughter aliquots by a combination of their isolation using CD31 and detection of miR-21 expression. These exosomes were confirmed to be not related to fragmented cells from testing for beta-tubulin. A global/discovery-based approach using mass spectrometry provided insights into early characterization of metabolomic profiles present in these tumor cells. Ample amounts of high quality DNA (226 ng/ul concentrations; 11.3 ug total) were recovered from the dislodged, excess cells in the wash for molecular studies. Finally, from viable cells recovered in one of the daughter wash aliquots, the ability to grow organoids was proven to be possible and reproducible. DISCUSSION/SIGNIFICANCE: Based on these results, the value of the clinical specimen can be markedly expanded for utilization in research and possible clinical use without detracting from the parent tissue. This non-destructive, easy to adopt wash procedure can potentially lead to an influx of data that may ultimately prove useful in improving patient care. |
| first_indexed | 2024-04-09T16:16:06Z |
| format | Article |
| id | doaj.art-81d638abdbac44b9b95ec6717202c633 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-09T16:16:06Z |
| publishDate | 2023-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-81d638abdbac44b9b95ec6717202c6332023-04-24T05:55:55ZengCambridge University PressJournal of Clinical and Translational Science2059-86612023-04-01710610710.1017/cts.2023.400360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimensWilfrido Mojica0Bei Yang1Chang Chieh Hsu2Yun Wu3Alexandra Izydorczak4Troy D. Wood5Dara Cho6Natesh Parashurama7Donald Yergeau8Supriya D Mahajan9State University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloState University of New York, BuffaloUniversity at Buffalo, SUNYOBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can contribute to the further development of Precision Medicine. METHODS/STUDY POPULATION: Transurethral resection of bladder tumors were selected for testing based on availability and tissue composition. A wash step was used to generate daughter aliquots composed of dislodged cells and a solution with prior contact to the parent tissue. This wash step served two purposes: 1) reduce the amount of contaminating material from spreading to other cases, a problem known to be associated with this type of specimen; and 2) create aliquots from which additional informative data could be generated. These daughter aliquots were then examined to determine their value as a source for exosome profiling, metabolomic studies, molecular characterization and organoid development. The parent tissue was not compromised, was able to undergo conventional processing and yielded results equivalent to unwashed specimens. RESULTS/ANTICIPATED RESULTS: Exosomes secreted by the tumor cells were identified to be present in the daughter aliquots by a combination of their isolation using CD31 and detection of miR-21 expression. These exosomes were confirmed to be not related to fragmented cells from testing for beta-tubulin. A global/discovery-based approach using mass spectrometry provided insights into early characterization of metabolomic profiles present in these tumor cells. Ample amounts of high quality DNA (226 ng/ul concentrations; 11.3 ug total) were recovered from the dislodged, excess cells in the wash for molecular studies. Finally, from viable cells recovered in one of the daughter wash aliquots, the ability to grow organoids was proven to be possible and reproducible. DISCUSSION/SIGNIFICANCE: Based on these results, the value of the clinical specimen can be markedly expanded for utilization in research and possible clinical use without detracting from the parent tissue. This non-destructive, easy to adopt wash procedure can potentially lead to an influx of data that may ultimately prove useful in improving patient care.https://www.cambridge.org/core/product/identifier/S2059866123004004/type/journal_article |
| spellingShingle | Wilfrido Mojica Bei Yang Chang Chieh Hsu Yun Wu Alexandra Izydorczak Troy D. Wood Dara Cho Natesh Parashurama Donald Yergeau Supriya D Mahajan 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens Journal of Clinical and Translational Science |
| title | 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens |
| title_full | 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens |
| title_fullStr | 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens |
| title_full_unstemmed | 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens |
| title_short | 360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens |
| title_sort | 360 waste not test more innovations in tissue processing to expand the testing of clinical specimens |
| url | https://www.cambridge.org/core/product/identifier/S2059866123004004/type/journal_article |
| work_keys_str_mv | AT wilfridomojica 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT beiyang 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT changchiehhsu 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT yunwu 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT alexandraizydorczak 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT troydwood 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT daracho 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT nateshparashurama 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT donaldyergeau 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens AT supriyadmahajan 360wastenottestmoreinnovationsintissueprocessingtoexpandthetestingofclinicalspecimens |