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European Journal of Applied Sciences – Vol. 12, No. 3

Publication Date: June 25, 2024

DOI:10.14738/aivp.123.17175.

Dackouo, B., Arama, D. P., Dembele, O., Toure, H. A., Mariko, M., Koumare, B. Y., & Kanoute, G. (2024). Overview in Abnormal

Read Cell Lipidomic Profiling for Human Chronic Disorder Diagnosis. European Journal of Applied Sciences, Vol - 12(3). 563-569.

Services for Science and Education – United Kingdom

Overview in Abnormal Read Cell Lipidomic Profiling for Human

Chronic Disorder Diagnosis

Blaise Dackouo

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Dominique Patomo Arama

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Ousmane Dembele

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Hamadoun Abba Toure

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Madani Mariko

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Benoît Yaranga Koumare

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

Gaoussou Kanoute

Faculty of Pharmacy (FAPH)/University of Sciences,

Technics and Technologies of Bamako (USTTB). P. O. Box 1805 Bamako, Mali

ABSTRACT

Red blood cell lipidomics has merged as new way to explore chronic disease

biomarkers. The aim of this review to explore ethics sample collection and

treatment for lipidomics purpose. Samples used to perform lipidomics is also

addressed herein. As a result, remarkable efforts are made is the field.

Keywords: Abnormal erythrocytes, Lipidomic, LC-MS

INTRODUCTION

OMICS approach is usable for red blood cell lipids profiling [1]. The exploration of lipids in red

cells to diagnose chronical diseases is common nowadays [12]. Sphingomyelin, ceramide, and

sphingosine are sphingolipids are explored in erythrocytes to diagnose lipid metabolism

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pathologies [13]. Changes in erythrocyte metabolism can affect their susceptibility to

hemolysis in routine storage [6]. Sphingolipids are erythrocyte biomarkers of some innate

diseases of red blood cells [13]. Changes in sphingolipid metabolism predict abnormal red cell

diseases [14]. To dates, lipidomic is viewed as a field of systems level analysis [30].

Erythrocytes eliminate wastes by vesiculation, a process disturbed in sickle cell disease,

glucose 6-phosphate dehydrogenase deficiency, spherocytosis, malaria, and other abnormal

red cell disorders [15]. In Glucose-6 phosphate dehydrogenase deficiency erythrocytes

haemolyze because of lipid oxidative insults making their storage less longer than normal

erythrocytes storage [5]. Active fatty acid desaturase of erythrocytes is sensitive to oxidant

stress, storage, duration, and alteration of the pyruvate/lactate ratio in G6PD deficient people

[4]. This change is known as oxidative stress [7]. Advances the field have shown that biofluids

as blood can be explored in incredible time [32]. Targeted lipidomic was adopted by the LIPID

MAPS consortium for the analysis of mammalian lipids [30]. Lipid profiles of chronic

disorders explore plasma, red cells or any cell [11].

To understand how to operate lipid extraction in red cell and performing metabolic profiling

require, the aim of this bibliographic search was survey recent work in lipidomic method.

MATERIALS AND METHOD

Materials

As any omics study, LC-MS is a key analytical method used in Lipidomic in addition, sample

preparation is an important factor in successful investigation [10]. Whole blood can be used

to explore red cell lipidomic [1]. Bone marrow is also usable [3]. Some reachers have

successfully worked on dry blood spots [33]. Human urines and plasma are also usable to

explore lipidomic in chronic pathologies [10]. Varied samples are usable to perform red cell

lipidomics.

Methods

Extracting Lipids for Lipidomic Purpose:

Lipidomic investigations can require whole blood samples collection in K3EDTA tubes [1].

They are centrifugated to sediment red cells so that plasma can be removed. Erythrocytes are

finally washed four times with physiological NaCl solution to get rid of white blood cells [1].

The washed red cells are kept in saline-adenine-glucose-mannitol or in a phosphate buffer

solution with a pH of 7.4 at 4° C [1]. Lipids are commonly extracted by a chloroform and

methanol-based protocol of Folch or Bligh and Dyer [30]. It is also possible to use pyridine to

effectively extract sphingolipids and phosphatidylcholine from intact S. cerevisiae [30].

Lipidomic exploration require special sampling and extraction methods for repeatability

purpose [7]. Another method called methyl-tert-butyl ether extraction has recently merged

with advantageous of no contamination [30].

Lipid Analytical Profiling:

Biomarker identification studies sometimes use LC-MS even LC-MS/MS methods for biological

process explorations [9]. Eugene. P. Kennedy is the pioneer in cell lipid sensing by exploring

phospholipid biosynthesis, before that whole lipids were addressed by Low resolution thin

layer chromatography [30]. This last approach is still used because easy and cheap [30]. FT-IR

is usable for the same purpose [1]. Gaz chromatography can separate of fatty acids from

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Dackouo, B., Arama, D. P., Dembele, O., Toure, H. A., Mariko, M., Koumare, B. Y., & Kanoute, G. (2024). Overview in Abnormal Read Cell Lipidomic

Profiling for Human Chronic Disorder Diagnosis. European Journal of Applied Sciences, Vol - 12(3). 563-569.

URL: http://dx.doi.org/10.14738/aivp.123.17175

simple mixtures however lipid detection and quantification in complex mixtures remains a

challenge [30]. GC-MS is successfully used [12]. Sphingolipids are analysed by mass

spectrometry after a step of mild alkaline hydrolysis to reduce ion suppression by other lipids

such as glycerophospholipids. ESI is the lipid ionization process of MS [30]. UPLC-HRMS as

well [15]. LC-MS and UPLC-MS with appropriate sample preparation are also usable [9].

Recently, rapid progresses have occurred in lipid research with the introduction of

electrospray ionization mass spectrometry and matrix assisted laser desorption ionization

mass spectrometry rendering cell lipidomic more possible [30]. LC-URMS is used for same

purpose [16]. [5]. LC-MS and UPLC-MS lipidomic performed on mouse bone marrow derived

macrophages following erythrophagocytosis confirmed lead to similar founding [3]. Gas

chromatography coupled with mass spectrometry has innovated lipid analysis, but their

assessment was not feasible at that time [30]. Recently, rapid progresses have occurred in

lipid research with the introduction of electrospray ionization mass spectrometry and matrix

assisted laser desorption ionization mass spectrometry rendering cell lipidomic more

possible [30]. In 2003 Han and Gross defined lipidomic as a global analysis of cell lipidomes

by a comprehensive MS approach [30].

Ethical

As any clinical study, lipidomic requires ethical comity approval because human blood

samples are collected from volunteers and the study must be conducted as guided by the

regulation of the Helsinki declaration in 1975 [1]. They must be conducted in according to the

guidelines of the Declaration of Helsinki, they must be approved by the Ethical Committee of

the Local Institution and informed consent must be obtained from all participatants [11].

RESULTS

Lipidomic of a class or a subclass is called a targeted lipidomic where different lipid classes

are analysed separately using extraction and analytical protocols designed specifically for

each lipid class [30]. Many chronic desorders are associated with change in red cell

metabolism and the susceptibility to hemolysis during a routine storage [6]. In glucosis six

phosphate dehydrogenase deficiency, erythrocytes remove damaged cell componens as

oxidized hemoglobin as well as damaged membrane constituents by microvesicles they

generate to prolong their lifespan [16]. Hemoglobin damage and altered phosphorylation of

membrane proteins as band-3 weaken the linkage between lipid bilayers and cell

cytoskeleton, resulting in membrane budding to microparticules [16]. Abnormal

hematological biomarkers have pathological and nutritionnal impacts [17]. Plasma and red

cell lipid metabolic profiling has supported that evidence in sickle cell disease anaemia [18].

People with this genetic desorder need appropriate advices and multidisciplinary cares [19].

Special treatments are suggested to prevent complications [20]. Further investigations are

required to better understand the mechanism of lipid alteration in red blood cell and discover

adequate treatments [21, 22]. In phospholipids, phosphatidylcholines and sphingomyelins

tend be more abundant in red cell outer leaflet [23]. Lipid metabolism profiling is possible on

other blood cells as blood mononuclear cells [24]. Recent studies predict great advances in the

field of inflammatory affections [25, 26]. Studies revealed an association of sickle disease with

adverse pregnancy outcomes, as maternas and neonatal anaemia, low birth weight, and

increased risk of stillbirth [27]. Metabolic markers of hemolysis and glucose 6-phosphate

dehydrogenasedeficiency are also associated with extremes in oxidative hemolysis and

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dysregulation in nicotinamide adenine dinucleotide phosphate and glutathione - dependent

detoxification pathways of oxidized lipids [28]. Lipid homeostasis is crucial in the

understanding of cell physiology, in specific lipid functions mechanisme and in that of the

local environment created [29]. Lipid metabolism disorders, as hypocholesterolemia and

hypertriglyceridemia, are associated with clinical events observed in sickle cell aneamia as a

result lipid are crucial in this multifactorial pathogenesis [31]. Many methods are used in lipid

metabolism profiling precisely GC-MS and LC-MS [32].

Figure 1: Chemical structures of sphingolipid and phospholipids [34].

DISCUSSION

The word lipidome appeared in 2001 and was used to describe a complete lipid contain of a

cell, tissue, or organism [30]. In mammalian, biomarkers are sometimes detectable by

sophisticated analytical methods; lipidomic is not an exception because LC-M or CG-MS are

commonly used for this purpose [9]. Same method was used to assess the impact of

hydroquinone and is metabolites on hepatic stellate cell. The study showed that

hydroquinone can reduce collagen synthesis slow the progress of liver fibrosis [10]. Lipid

class involved in the pathology occurrence can be established by the study data using

appropriate method [11]. Sphingomyelin, Ceramide and Shingosine are erythrocyte lipids

pointed to be associated with abnormal red cell chronic pothogies provided that their

metabolizing enzyme and changes can be observed in corpuscules and in plasma as well [12].

Some scientists suggest that red cells endure oxidative stress metabolic processes [13].

Functional lipidomic was introduced by Rilfors and Lindblom who coined it to describe the

study of role played by membrane lipids [30]. It is therefore a need to a perform farther

investigation to understand red cell physiology.

CONCLUSION

Lipidomic profiling is feasible on dry blood spots, whole blood, serum plasma and red blood

cells. The best mode of sample treatment is online method, gas chromatography and liquid

chromatography coupled with mass spectrometry are analytical methods that are used to

perform the job. These methods are usable to do targeted lipidomic. Human lipidomic as well

as any clinical study should abbey the guidelines of 1975 Helsinki declarations and study

volunteers accept to participate to the study by signing an informed consent form before a

sample is getting collected from them for the study.

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