Factors that influence the quality of RNA from the pancreas of organ donors

Tiffany Philips, Irina Kusmartseva, Ivan Gerling, Martha Campbell-Thompson, Clive Wasserfall, Alberto Pugliese, Jeffrey A. Longmate, Desmond A. Schatz, Mark A. Atkinson, John S. Kaddis

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Objectives: Attaining high-quality RNA fromthe tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, Methods: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN. Results: Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95%confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95% CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95% CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95% CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95% CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95% CI, 0.6-2.0; P < 0.01). Conclusions: A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA.

Original languageEnglish (US)
Pages (from-to)252-259
Number of pages8
JournalPancreas
Volume46
Issue number2
DOIs
StatePublished - Jan 1 2017

Fingerprint

Pancreas
Tissue Donors
RNA
Confidence Intervals
Odds Ratio
Laser Capture Microdissection
Lipase
Cause of Death
Autopsy
Hospitalization
Logistic Models
Biopsy
Research

All Science Journal Classification (ASJC) codes

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

Cite this

Philips, T., Kusmartseva, I., Gerling, I., Campbell-Thompson, M., Wasserfall, C., Pugliese, A., ... Kaddis, J. S. (2017). Factors that influence the quality of RNA from the pancreas of organ donors. Pancreas, 46(2), 252-259. https://doi.org/10.1097/MPA.0000000000000717

Factors that influence the quality of RNA from the pancreas of organ donors. / Philips, Tiffany; Kusmartseva, Irina; Gerling, Ivan; Campbell-Thompson, Martha; Wasserfall, Clive; Pugliese, Alberto; Longmate, Jeffrey A.; Schatz, Desmond A.; Atkinson, Mark A.; Kaddis, John S.

In: Pancreas, Vol. 46, No. 2, 01.01.2017, p. 252-259.

Research output: Contribution to journalArticle

Philips, T, Kusmartseva, I, Gerling, I, Campbell-Thompson, M, Wasserfall, C, Pugliese, A, Longmate, JA, Schatz, DA, Atkinson, MA & Kaddis, JS 2017, 'Factors that influence the quality of RNA from the pancreas of organ donors', Pancreas, vol. 46, no. 2, pp. 252-259. https://doi.org/10.1097/MPA.0000000000000717
Philips T, Kusmartseva I, Gerling I, Campbell-Thompson M, Wasserfall C, Pugliese A et al. Factors that influence the quality of RNA from the pancreas of organ donors. Pancreas. 2017 Jan 1;46(2):252-259. https://doi.org/10.1097/MPA.0000000000000717
Philips, Tiffany ; Kusmartseva, Irina ; Gerling, Ivan ; Campbell-Thompson, Martha ; Wasserfall, Clive ; Pugliese, Alberto ; Longmate, Jeffrey A. ; Schatz, Desmond A. ; Atkinson, Mark A. ; Kaddis, John S. / Factors that influence the quality of RNA from the pancreas of organ donors. In: Pancreas. 2017 ; Vol. 46, No. 2. pp. 252-259.
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abstract = "Objectives: Attaining high-quality RNA fromthe tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, Methods: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN. Results: Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95{\%}confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95{\%} CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95{\%} CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95{\%} CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95{\%} CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95{\%} CI, 0.6-2.0; P < 0.01). Conclusions: A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA.",
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AU - Philips, Tiffany

AU - Kusmartseva, Irina

AU - Gerling, Ivan

AU - Campbell-Thompson, Martha

AU - Wasserfall, Clive

AU - Pugliese, Alberto

AU - Longmate, Jeffrey A.

AU - Schatz, Desmond A.

AU - Atkinson, Mark A.

AU - Kaddis, John S.

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N2 - Objectives: Attaining high-quality RNA fromthe tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, Methods: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN. Results: Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95%confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95% CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95% CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95% CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95% CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95% CI, 0.6-2.0; P < 0.01). Conclusions: A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA.

AB - Objectives: Attaining high-quality RNA fromthe tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, Methods: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN. Results: Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95%confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95% CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95% CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95% CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95% CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95% CI, 0.6-2.0; P < 0.01). Conclusions: A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA.

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