Postmortem mRNA profiling II: Practical considerations

Vennemann, Marielle; Koppelkamm, Antje

doi:10.1016/j.forsciint.2010.07.007

Cited by 26 publications

(22 citation statements)

References 39 publications

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“…Five commonly used reference genes were chosen ( Actb , Gapdh , Hprt , Cyp2E1 and Ppia ) as widely accepted stable expressed genes [16], [17], [38]. In spite of the original thought of an ubiquitous expression of such genes, several studies already found an existing variation in the expression of commonly used reference genes [25], [36], [39].…”

Section: Discussionmentioning

confidence: 99%

Profiling of RNA Degradation for Estimation of Post Morterm Interval

et al. 2013

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An estimation of the post mortem interval (PMI) is frequently touted as the Holy Grail of forensic pathology. During the first hours after death, PMI estimation is dependent on the rate of physical observable modifications including algor, rigor and livor mortis. However, these assessment methods are still largely unreliable and inaccurate. Alternatively, RNA has been put forward as a valuable tool in forensic pathology, namely to identify body fluids, estimate the age of biological stains and to study the mechanism of death. Nevertheless, the attempts to find correlation between RNA degradation and PMI have been unsuccessful. The aim of this study was to characterize the RNA degradation in different post mortem tissues in order to develop a mathematical model that can be used as coadjuvant method for a more accurate PMI determination. For this purpose, we performed an eleven-hour kinetic analysis of total extracted RNA from murine's visceral and muscle tissues. The degradation profile of total RNA and the expression levels of several reference genes were analyzed by quantitative real-time PCR. A quantitative analysis of normalized transcript levels on the former tissues allowed the identification of four quadriceps muscle genes (Actb, Gapdh, Ppia and Srp72) that were found to significantly correlate with PMI. These results allowed us to develop a mathematical model with predictive value for estimation of the PMI (confidence interval of ±51 minutes at 95%) that can become an important complementary tool for traditional methods.

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Section: Discussionmentioning

confidence: 99%

Profiling of RNA Degradation for Estimation of Post Morterm Interval

et al. 2013

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“…Postmortem intervals and conditions as well as antemortem factors such as age, gender, body mass, and the physiological/pathological conditions of the agonal phase can profoundly affect expression profiles and half-lives of certain gene transcripts in postmortem tissues, obscuring the employment of molecular biomarkers to investigate the cause and process of death (Durrenberger et al, 2010;Preece and Cairns, 2003;Tomita et al, 2004). Therefore, data normalization and RNA integrity have been essential issues in the field of forensic molecular pathology (Vennemann and Koppelkamm, 2010b). Various normalization methods using the total amount of tissue, DNA, or RNA, and endogenous reference genes have been proposed (Huggett et al, 2005;Talaat et al, 2002;Vandesompele et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Quantitative Analyses of Postmortem Heat Shock Protein mRNA Profiles in the Occipital Lobes of Human Cerebral Cortices: Implications in Cause of Death

Chung

Seo

Kim

et al. 2012

Molecules and Cells

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Quantitative RNA analyses of autopsy materials to diagnose the cause and mechanism of death are challenging tasks in the field of forensic molecular pathology. Alterations in mRNA profiles can be induced by cellular stress responses during supravital reactions as well as by lethal insults at the time of death. Here, we demonstrate that several gene transcripts encoding heat shock proteins (HSPs), a gene family primarily responsible for cellular stress responses, can be differentially expressed in the occipital region of postmortem human cerebral cortices with regard to the cause of death. HSPA2 mRNA levels were higher in subjects who died due to mechanical asphyxiation (ASP), compared with those who died by traumatic injury (TI). By contrast, HSPA7 and A13 gene transcripts were much higher in the TI group than in the ASP and sudden cardiac death (SCD) groups. More importantly, relative abundances between such HSP mRNA species exhibit a stronger correlation to, and thus provide more discriminative information on, the death process than does routine normalization to a housekeeping gene. Therefore, the present study proposes alterations in HSP mRNA composition in the occipital lobe as potential forensic biological markers, which may implicate the cause and process of death.

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“…Multiple classes of RNA species such as messenger RNAs (mRNAs), ribosomal RNAs (rRNAs), and microRNAs have been tested for this purpose. However, correlating the degradation of RNA species with PMI still remains an area of concern, primarily owing to difficulties in the standardization and/or normalization of quantitative analyses with sufficient accuracy and reproducibility (Vennemann and Koppelkamm 2010a; 2010b). Postmortem RNA profiles represent preexisting pathophysiological states along with postmortem conditions, thereby leading to large variations among individuals.…”

Section: Introductionmentioning

confidence: 99%

Cell Death-Associated Ribosomal RNA Cleavage in Postmortem Tissues and Its Forensic Applications

Kim

Kyeong

et al. 2017

Molecules and Cells

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Estimation of postmortem interval (PMI) is a key issue in the field of forensic pathology. With the availability of quantitative analysis of RNA levels in postmortem tissues, several studies have assessed the postmortem degradation of constitutively expressed RNA species to estimate PMI. However, conventional RNA quantification as well as biochemical and physiological changes employed thus far have limitations related to standardization or normalization. The present study focuses on an interesting feature of the subdomains of certain RNA species, in which they are site-specifically cleaved during apoptotic cell death. We found that the D8 divergent domain of ribosomal RNA (rRNA) bearing cell death-related cleavage sites was rapidly removed during postmortem RNA degradation. In contrast to the fragile domain, the 5′ terminal region of 28S rRNA was remarkably stable during the postmortem period. Importantly, the differences in the degradation rates between the two domains in mammalian 28S rRNA were highly proportional to increasing PMI with a significant linear correlation observed in mice as well as human autopsy tissues. In conclusion, we demonstrate that comparison of the degradation rates between domains of a single RNA species provides quantitative information on postmortem degradation states, which can be applied for the estimation of PMI.

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Postmortem mRNA profiling II: Practical considerations

Cited by 26 publications

References 39 publications

Profiling of RNA Degradation for Estimation of Post Morterm Interval

Profiling of RNA Degradation for Estimation of Post Morterm Interval

Quantitative Analyses of Postmortem Heat Shock Protein mRNA Profiles in the Occipital Lobes of Human Cerebral Cortices: Implications in Cause of Death

Cell Death-Associated Ribosomal RNA Cleavage in Postmortem Tissues and Its Forensic Applications

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