Top

Published in:

29-09-2022 | Biomarkers | Original Article

Candidate biomarkers in brown adipose tissue for post-mortem diagnosis of fatal hypothermia

Authors: Miao Zhang, Ning Wang, Xiang-Shen Guo, Lin-Lin Wang, Peng-Fei Wang, Zhi-Peng Cao, Fu-Yuan Zhang, Zi-Wei Wang, Da-Wei Guan, Rui Zhao

Published in: International Journal of Legal Medicine | Issue 1/2024

Abstract

Post-mortem diagnosis of fatal hypothermia (FHT) is challenging in forensic practice because traditional morphological and biochemical methods lack specificity. Recent studies have reported that brown adipose tissue (BAT) is activated during cold-induced non-shivering thermogenesis in mammals, but BAT has not been used to diagnose FHT. The aim of this study was to identify novel biomarkers in BAT for FHT based on morphological changes and differential protein expression. Two FHT animal models were created by exposing mice to 4 or −20 °C at 50% humidity. Morphologically, the unilocular lipid droplet content was significantly increased in BAT of FHT model mice compared with that of control mice. Proteomics analysis revealed a total of 283 and 266 differentially expressed proteins (DEPs) between the 4 or −20 °C FHT subgroups and control group, respectively. In addition, 140 proteins were shared between the FHT subgroups. GO and KEGG analyses revealed that the shared DEPs were mainly enriched in pathways associated with metabolism, oxidative phosphorylation, and thermogenesis. Further screening (|log₂FC| > 1.6, q-value (FDR) < 0.05) identified GMFB, KDM1A, DDX6, RAB1B, SHMT-1, CLPTM1, and LMF1 as candidate biomarkers of FHT. Subsequent validation experiments were performed in FHT model mice using classic immunohistochemistry and western blotting. RAB1B and GMFB expression was further verified in BAT specimens from human cases of FHT. The results demonstrate that BAT can be used as a target organ for FHT diagnosis employing RAB1B and GMFB as biological markers, thus providing a new strategy for the post-mortem diagnosis of FHT in forensic practice.

Available only for authorised users

Hajat S, Kovats RS, Lachowycz K (2007) Heat-related and cold-related deaths in England and Wales: who is at risk? Occup Environ Med 64:93–100. https://doi.org/10.1136/oem.2006.029017CrossRefPubMed

Berko J, Ingram DD, Saha S, Parker JD (2014) Deaths attributed to heat, cold, and other weather events in the United States, 2006-2010. Natl Health Stat Rep 1-15

Herity B, Daly L, Bourke GJ, Horgan JM (1991) Hypothermia and mortality and morbidity. An epidemiological analysis. J Epidemiol Community Health 45:19. https://doi.org/10.1136/jech.45.1.19CrossRefPubMedPubMedCentral

Lin H, Deng K, Zhang J et al (2019) Biochemical detection of fatal hypothermia and hyperthermia in affected rat hypothalamus tissues by Fourier transform infrared spectroscopy. Biosci Rep 39. https://doi.org/10.1042/bsr20181633

Palmiere C, Teresinski G, Hejna P (2014) Postmortem diagnosis of hypothermia. Int J Legal Med 128:607–614. https://doi.org/10.1007/s00414-014-0977-1CrossRefPubMed

Paal P, Rauch S (2018) Indoor accidental hypothermia in the elderly: an emerging lethal entity in the 21st century. Emerg Med J 35:667–668. https://doi.org/10.1136/emermed-2018-207804CrossRefPubMed

Charkoudian N (2003) Skin blood flow in adult human thermoregulation: how it works, when it does not, and why. Mayo Clin Proc 78:603–612. https://doi.org/10.4065/78.5.603CrossRefPubMed

Ouellet V, Labbe SM, Blondin DP et al (2012) Brown adipose tissue oxidative metabolism contributes to energy expenditure during acute cold exposure in humans. J Clin Invest 122:545–552. https://doi.org/10.1172/jci60433CrossRefPubMedPubMedCentral

Cannon B, Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84:277–359. https://doi.org/10.1152/physrev.00015.2003CrossRefPubMed

10.

Richard D, Picard F (2011) Brown fat biology and thermogenesis. Front Biosci (Landmark Ed) 16:1233–1260. https://doi.org/10.2741/3786CrossRefPubMed

11.

Morrison SF, Madden CJ (2014) Central nervous system regulation of brown adipose tissue. Compr Physiol 4:1677–1713. https://doi.org/10.1002/cphy.c140013CrossRefPubMedPubMedCentral

12.

Bal NC, Singh S, Reis FCG et al (2017) Both brown adipose tissue and skeletal muscle thermogenesis processes are activated during mild to severe cold adaptation in mice. J Biol Chem 292:16616–16625. https://doi.org/10.1074/jbc.M117.790451CrossRefPubMedPubMedCentral

13.

Lim S, Honek J, Xue Y et al (2012) Cold-induced activation of brown adipose tissue and adipose angiogenesis in mice. Nat Protoc 7:606–615. https://doi.org/10.1038/nprot.2012.013CrossRefPubMed

14.

Brychta RJ, Chen KY (2017) Cold-induced thermogenesis in humans. Eur J Clin Nutr 71:345–352. https://doi.org/10.1038/ejcn.2016.223CrossRefPubMed

15.

van Marken Lichtenbelt WD, Vanhommerig JW, Smulders NM et al (2009) Cold-activated brown adipose tissue in healthy men. N Engl J Med 360:1500–1508. https://doi.org/10.1056/NEJMoa0808718CrossRefPubMed

16.

Nedergaard J, Golozoubova V, Matthias A, Asadi A, Jacobsson A, Cannon B (2001) UCP1: the only protein able to mediate adaptive non-shivering thermogenesis and metabolic inefficiency. Biochim Biophys Acta 1504:82–106. https://doi.org/10.1016/s0005-2728(00)00247-4CrossRefPubMed

17.

Green HJ, Burnett M, Kollias H, Ouyang J, Smith I, Tupling S (2012) Can increases in capillarization explain the early adaptations in metabolic regulation in human muscle to short-term training? Can J Physiol Pharmacol 90:557–566. https://doi.org/10.1139/y2012-013CrossRefPubMed

18.

Wang N, Lu HY, Li X et al (2019) ZW290 increases cold tolerance by inducing thermogenesis via the upregulation of uncoupling protein 1 in brown adipose tissue in vitro and in vivo. Lipids 54:265–276. https://doi.org/10.1002/lipd.12148CrossRefPubMed

19.

Bright F, Winskog C, Walker M, Byard RW (2013) Why are Wischnewski spots not always present in lethal hypothermia? The results of testing a stress-reduced animal model. J Forensic Legal Med 20:785–787. https://doi.org/10.1016/j.jflm.2013.05.003CrossRef

20.

Tsokos M, Rothschild MA, Madea B, Rie M, Sperhake JP (2006) Histological and immunohistochemical study of Wischnewsky spots in fatal hypothermia. Am J Forensic Med Pathol 27:70–74. https://doi.org/10.1097/01.paf.0000202716.06378.91CrossRefPubMed

21.

Lin H, Zou D, Luo Y et al (2019) Postmortem diagnosis of fatal hypothermia/hyperthermia by spectrochemical analysis of plasma. Forensic Sci Med Pathol 15:332–341. https://doi.org/10.1007/s12024-019-00111-8CrossRefPubMed

22.

Palmiere C, Bardy D, Letovanec I et al (2013) Biochemical markers of fatal hypothermia. Forensic Sci Int 226:54–61. https://doi.org/10.1016/j.forsciint.2012.12.007CrossRefPubMed

23.

Lin H, Guo X, Luo Y et al (2020) Postmortem diagnosis of fatal hypothermia by Fourier transform infrared spectroscopic analysis of edema fluid in formalin-fixed, paraffin-embedded lung tissues. J Forensic Sci 65:846–854. https://doi.org/10.1111/1556-4029.14260CrossRefPubMed

24.

Rousseau G, Chao de la Barca JM, Rougé-Maillart C et al (2021) Preliminary metabolomic profiling of the vitreous humor from hypothermia fatalities. J Proteome Res 20:2390–2396. https://doi.org/10.1021/acs.jproteome.0c00901CrossRefPubMed

25.

Rousseau G, Chao de la Barca JM, Rougé-Maillart C et al (2019) A serum metabolomics signature of hypothermia fatalities involving arginase activity, tryptophan content, and phosphatidylcholine saturation. Int J Legal Med 133:889–898. https://doi.org/10.1007/s00414-018-1937-yCrossRefPubMed

26.

Sjöström L, Björntorp P, Vrána J (1971) Microscopic fat cell size measurements on frozen-cut adipose tissue in comparison with automatic determinations of osmium-fixed fat cells. J Lipid Res 12:521–530CrossRefPubMed

27.

Bartelt A, Bruns OT, Reimer R et al (2011) Brown adipose tissue activity controls triglyceride clearance. Nat Med 17:200–205. https://doi.org/10.1038/nm.2297CrossRefPubMed

28.

Valgas P, da Silva C, Hernández-Saavedra D, White JD, Stanford KI (2019) Cold and exercise: therapeutic tools to activate brown adipose tissue and combat obesity. Biology (Basel) 8:9. https://doi.org/10.3390/biology8010009CrossRef

29.

Saito M, Okamatsu-Ogura Y, Matsushita M et al (2009) High incidence of metabolically active brown adipose tissue in healthy adult humans: effects of cold exposure and adiposity. Diabetes 58:1526–1531. https://doi.org/10.2337/db09-0530CrossRefPubMedPubMedCentral

30.

McDonald RB, Horwitz BA (1999) Brown adipose tissue thermogenesis during aging and senescence. J Bioenerg Biomembr 31:507–516. https://doi.org/10.1023/a:1005404708710CrossRefPubMed

31.

Kotzbeck P, Giordano A, Mondini E et al (2018) Brown adipose tissue whitening leads to brown adipocyte death and adipose tissue inflammation. J Lipid Res 59:784–794. https://doi.org/10.1194/jlr.M079665CrossRefPubMedPubMedCentral

32.

Li J, Smith LS, Zhu HJ (2021) Data-independent acquisition (DIA): an emerging proteomics technology for analysis of drug-metabolizing enzymes and transporters. Drug Discov Today Technol 39:49–56. https://doi.org/10.1016/j.ddtec.2021.06.006CrossRefPubMedPubMedCentral

33.

Shephard RJ (1993) Metabolic adaptations to exercise in the cold. An update. Sports Med 16:266–289. https://doi.org/10.2165/00007256-199316040-00005CrossRefPubMed

34.

Prieto JA, Estruch F, Córcoles-Sáez I et al (2020) Pho85 and PI(4,5)P(2) regulate different lipid metabolic pathways in response to cold. Biochim Biophys Acta Mol Cell Biol Lipids 1865:158557. https://doi.org/10.1016/j.bbalip.2019.158557CrossRefPubMed

35.

Hutagalung AH, Novick PJ (2011) Role of Rab GTPases in membrane traffic and cell physiology. Physiol Rev 91:119–149. https://doi.org/10.1152/physrev.00059.2009CrossRefPubMed

36.

Romero N, Dumur CI, Martinez H et al (2013) Rab1b overexpression modifies Golgi size and gene expression in HeLa cells and modulates the thyrotrophin response in thyroid cells in culture. Mol Biol Cell 24:617–632. https://doi.org/10.1091/mbc.E12-07-0530CrossRefPubMedPubMedCentral

37.

Gohlke S, Zagoriy V, Cuadros Inostroza A et al (2019) Identification of functional lipid metabolism biomarkers of brown adipose tissue aging. Mol Metab 24:1–17. https://doi.org/10.1016/j.molmet.2019.03.011CrossRefPubMedPubMedCentral

38.

Wang BR, Zaheer A, Lim R (1992) Polyclonal antibody localizes glia maturation factor beta-like immunoreactivity in neurons and glia. Brain Res 591:1–7. https://doi.org/10.1016/0006-8993(92)90971-bCrossRefPubMed

39.

Zaheer A, Fink BD, Lim R (1993) Expression of glia maturation factor beta mRNA and protein in rat organs and cells. J Neurochem 60:914–920. https://doi.org/10.1111/j.1471-4159.1993.tb03237.xCrossRefPubMed

40.

Zaheer A, Mathur SN, Lim R (2002) Overexpression of glia maturation factor in astrocytes leads to immune activation of microglia through secretion of granulocyte-macrophage-colony stimulating factor. Biochem Biophys Res Commun 294:238–244. https://doi.org/10.1016/s0006-291x(02)00467-9CrossRefPubMed

41.

Hotta N, Aoyama M, Inagaki M et al (2005) Expression of glia maturation factor beta after cryogenic brain injury. Brain Res Mol Brain Res 133:71–77. https://doi.org/10.1016/j.molbrainres.2004.09.027CrossRefPubMed

42.

Diestel A, Troeller S, Billecke N, Sauer IM, Berger F, Schmitt KR (2010) Mechanisms of hypothermia-induced cell protection mediated by microglial cells in vitro. Eur J Neurosci 31:779–787. https://doi.org/10.1111/j.1460-9568.2010.07128.xCrossRefPubMed

43.

Teresiński G, Buszewicz G, Madro R (2005) Biochemical background of ethanol-induced cold susceptibility. Leg Med (Tokyo) 7:15–23. https://doi.org/10.1016/j.legalmed.2004.07.002CrossRefPubMed

44.

Lee YH, Mottillo EP, Granneman JG (2014) Adipose tissue plasticity from WAT to BAT and in between. Biochim Biophys Acta 1842:358–369. https://doi.org/10.1016/j.bbadis.2013.05.011CrossRefPubMed

Title: Candidate biomarkers in brown adipose tissue for post-mortem diagnosis of fatal hypothermia
Authors: Miao Zhang
Ning Wang
Xiang-Shen Guo
Lin-Lin Wang
Peng-Fei Wang
Zhi-Peng Cao
Fu-Yuan Zhang
Zi-Wei Wang
Da-Wei Guan
Rui Zhao
Publication date: 29-09-2022
Publisher: Springer Berlin Heidelberg
Keyword: Biomarkers
Published in: International Journal of Legal Medicine / Issue 1/2024
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI: https://doi.org/10.1007/s00414-022-02897-9

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Candidate biomarkers in brown adipose tissue for post-mortem diagnosis of fatal hypothermia

Abstract

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2024

A sudden death, an aortic rupture, and an unexpected cause: a report about suspected child abuse

Computer-aided craniofacial superimposition validation study: the identification of the leaders and participants of the Polish-Lithuanian January Uprising (1863–1864)

How to conceive the dignity of the dead? A dispositional account

Proteomic analysis discovers potential biomarkers of early traumatic axonal injury in the brainstem

Caught in the act: impact of Crematogaster cf. liengmei (Hymenoptera: Formicidae) necrophagous behavior on neonate pigs (Sus scrofa domesticus L.) in the Western Cape Province of South Africa

Sr–Pb isotope differences in pre- and post-burial human bone, teeth, and hair keratin: implications for isotope forensics