Assessment of chylomicronemia using vitamin e

Abstract

Described are methods related to the use of vitamin E as a clinical marker in subjects suffering from severe hypertriglyceridemia. One particular method comprises measuring a level of vitamin E in a subject's blood suffering from severe hypertriglyceridemia to assess a blood level of chylomicrons-related triglycerides. Another particular method comprises measuring a level of vitamin E in a subject's blood suffering from severe hypertriglyceridemia to distinguish between triglycerides and lipoproteins issued from the exogenous pathway versus those of the endogenous pathway. Another particular method comprises measuring vitamin E in a subject suffering from severe hypertriglyceridemia for assessing whether hypolipidemic treatments are effective or not, and to which extent. Another particular method is for assessing a risk of acute pancreatitis in a subject suffering from hypertriglyceridemia. Also described is a method for assessing vitamin E deficiency and kits comprising vitamin E.

Description

ASSESSMENT OF CHYLOMICRONEMIA USING VITAMIN ECROSS REFERENCE TO RELATED APPLICATION

[0001] This patent application claims priority to provisional patent application US 63 / 587,192 filed on October 02, 2023, the content of which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION

[0002] The invention relates to the field of lipids-related diseases, and more particularly to the use of vitamin E as a clinical marker in subjects suffering from severe hypertriglyceridemia.BACKGROUND OF THE INVENTION

[0003] Hypertriglyceridemia is characterized by the accumulation in the blood of lipoproteins rich in triglycerides (TG) originating from the exogenous (chylomicrons and remnants) or endogenous (VLDL and remnants) metabolic pathways. There are several forms of hypertriglyceridemia and the most severe form, hyperchylomicronemia, is defined by the blood accumulation of chylomicrons. Chylomicrons are large triglyceride-rich lipoproteins produced in enterocytes from dietary lipids — namely, fatty acids, and cholesterol. The chylomicrons are known to transport dietary fat after a meal. Chylomicronemia is therefore a postprandial metabolic disturbance characterized by a triglyceridemia greater than 10 mmol / L. The prevalence of chylomicronemia is estimated at 1 person out 600 (all causes) in the Western world. Chylomicronemia can be sustained or recurrent and several genetic, physiological, environmental factors or health conditions can favor the appearance of this condition. Chylomicronemia is considered to be sustained when the triglyceridemia is most often higher than 10 mmol / L and refractory to conventional hypolipidemic treatments.

[0004] Natural vitamin E includes four tocopherols and four tocotrienols. RRR-alpha- tocopherol is the most abundant form in nature and has the highest biological activity. Vitamin E is transported in blood lipoproteins. After its intestinal absorption vitamin E is packaged into chylomicrons, which along the lymphatic pathway are secreted into thesystemic circulation (Gagne A., et al., J Obstet Gynaecol Can, 2008;31 (3):210-217). Chylomicrons are known to transport vitamin E at their surface. By the action of lipoprotein lipase (LPL), part of the tocopherols transported in chylomicrons are taken up by extrahepatic tissues, and the remnant chylomicrons transport the remaining tocopherols to the liver (E Herrera and C Barbas, Vitamin E: action, metabolism and perspectives. J Physiol Biochem, 2001 Mar;57(1):43-56).

[0005] Despite the above, prior to the present invention, it had never been suggested to use vitamin E measurements as a clinical marker of a subject’s blood level of chylomicrons-related triglycerides.

[0006] Prior to the present invention, it had never been suggested either to measure vitamin E in the blood of a subject to clinically distinguish between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway.

[0007] Likewise, the present inventors are the first to teach measuring levels of vitamin E in a subject suffering from severe hypertriglyceridemia for assessing efficacy of hypolipidemic therapies.

[0008] Also, it had never been suggested prior to the present invention to assess the risks of acute pancreatitis in subjects suffering from severe hypertriglyceridemia by measuring a level of vitamin E in the subject’s blood.

[0009] Prior to the present invention, it had never been suggested either to assess a need in vitamin E in a subject suffering from severe hypertriglyceridemia by measuring a level of chylomicrons in the blood of the subject.

[0010] There is thus a need for new methods to assess chylomicrons-related triglycerides in subjects.

[0011] There is also a need for a better distinction between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway, particularly subjects suffering from hypertriglyceridemia.

[0012] There is also a need for assessing Vitamin E bioavailability in the blood, and not only chylomicron-bound vitamin E.

[0013] There is also a need for new methods to assess efficacy of hypolipidemic therapies.

[0014] There is also a need for proper assessment of subjects suffering from hypertriglyceridemia with respect to a risk of acute pancreatitis and other morbidities.

[0015] There is also a need for improved methods for treating a subject suffering from severe hypertriglyceridemia.

[0016] The present invention addresses these needs and other needs as it will be apparent from the review of the disclosure and description of the features of the invention hereinafter.BRIEF SUMMARY OF THE INVENTION

[0017] According to one aspect, the invention relates a method to assess a blood level of chylomicrons-related triglycerides in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood, wherein the level of vitamin E is indicative of the blood level of chylomicrons-related triglycerides.

[0018] According to another aspect, the invention relates to a method to distinguish between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway in blood of a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood, wherein the level of vitamin E is indicative of levels of triglycerides and lipoproteins issued from the exogenous pathway.

[0019] According to another aspect, the invention relates to a method to assess efficacy of an hypolipidemic therapy in a subject suffering from severe hypertriglyceridemia, the method comprising: (i) measuring a first level of vitamin E in thesubject’s blood prior to beginning of the hypolipidemic therapy; (ii) measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy.

[0020] According to another aspect, the invention relates to a method to assess a risk of acute pancreatitis in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood; wherein said level of vitamin E is indicative of the blood level of chylomicrons; and wherein a higher level of chylomicrons is indicative of a higher risk of acute pancreatitis for the subject.

[0021] According to another aspect, the invention relates to a method to assess a need of vitamin E in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of chylomicrons in the blood of the subject; wherein said level of chylomicrons is indicative of a level of vitamin E that is not bioavailable to the subject; and wherein the level of vitamin E that is not bioavailable is indicative of a need of vitamin E.

[0022] According to another aspect, the invention relates to a kit for assessing efficacy of an hypolipidemic therapy in a subject suffering from severe hypertriglyceridemia, the kit comprising multiple doses of vitamin E, and wherein said doses of vitamin E are comprised in a blister package, a capsule strip, a reclosable bottle, a powder pack.

[0023] According to another aspect, the invention relates to the use of a vitamin E blood measurement as a clinical marker of a subject’s blood level of chylomicrons-related triglycerides.

[0024] According to another aspect, the invention relates to the use of a vitamin E blood measurement to clinically distinguish in a subject between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway.

[0025] According to another aspect, the invention relates to the use of a vitamin E blood measurement in a subject suffering from severe hypertriglyceridemia for one or more of: (i) assessing efficacy of hypolipidemic therapies; assess the subject’s risks of an acute pancreatitis; and assess the subject’s need in vitamin E.

[0026] According to another aspect, the invention relates to a method for treating a subject suffering from severe hypertriglyceridemia, the method comprising:(i) measuring a first level of vitamin E in the subject’s blood prior to beginning of an hypolipidemic therapy;(ii) measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy; wherein said measurements of vitamin E provide a clinical marker of the subject’s blood level of chylomicrons-related triglycerides;(iii) providing a treatment plan taking into consideration the subject’s blood level of chylomicrons-related triglycerides.

[0027] Additional aspects, advantages and features of the present invention will become more apparent upon reading of the following non-restrictive description of preferred embodiments which are exemplary and should not be interpreted as limiting the scope of the invention.BRIEF DESCRIPTION OF THE FIGURES

[0028] NONEDETAILED DESCRIPTION OF EMBODIMENTS

[0029] In the following description of the embodiments, references to the accompanying figures are illustrations of an example by which the invention may be practised. It will be understood that other embodiments may be made without departing from the scope of the invention disclosed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skilled in the art to which the invention belongs.Overview

[0030] The invention relates to the field of lipids-related diseases. More particularly, the invention proposes using vitamin E level(s) in blood in subjects suffering from severe hypertriglyceridemia, as a clinical marker of a subject’s blood level of chylomicrons-related triglycerides. In accordance with the present invention using vitamin E level(s) in blood isalso useful for clinically distinguishing between triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway, for assessing efficacy of hypolipidemic therapies, for assessing a subject’s risks of an acute pancreatitis and / or for assessing a subject’s need in vitamin E, among possible applications.

[0031] As is known, Vitamin E is a group of eight fat soluble compounds that include four tocopherols and four tocotrienols. A worldwide summary of more than one hundred human studies reported a median of 22.1 pmol / L for serum a-tocopherol, and defined a-tocopherol deficiency as less than 12 pmol / L in adults. It cited a recommendation that serum a-tocopherol concentration be >30 pmol / L to optimize health benefits (Peter S, et al. (December 2015). "A Systematic Review of Global Alpha-Tocopherol Status as Assessed by Nutritional Intake Levels and Blood Serum Concentrations". International Journal for Vitamin and Nutrition Research, 85 (5-6): 261-281. doi: 10.1024 / 0300- 9831 / a000281. PMID 27414419).

[0032] It is also known that Vitamin E is absorbed and transported by chylomicrons. The present invention is based on the unexpected discovery that blood levels of vitamin E are higher than normal (2-10 times) in subjects suffering from a severe hypertriglyceridemia (e.g., chylomicronemia). Since the vitamin E is contained in the chylomicrons and cannot be released therefrom, it is not bioavailable. Therefore, it has been found also that reducing levels of chylomicrons also reduce levels of vitamin E. In contradistinction, the inventors knew that the blood levels of vitamin E are lower in subjects suffering from abetalipoproteinemia. And because the vitamin E is contained in the chylomicrons, these subjects thus require supplements of vitamin E.

[0033] As demonstrated hereinafter in the Examples, the present invention proposes using vitamin E as a clinical marker of chylomicrons-related triglycerides. According to the principles of the present invention, if the measured blood level of vitamin E is elevated in presence of sustained chylomicronemia, this elevated level suggests that the measured level of vitamin E mainly represents chylomicron-bound vitamin E. On the other hand, if and when chylomicrons decrease in size and / or if and when the level of chylomicronsdecreases, the vitamin E concentration decreases as well and better represent bio- available vitamin E.

[0034] As used herein, the term “subject” includes a human, more preferably a human patient in need of treatment. Even more preferably the subject is a human patient diagnosed or susceptible to suffer from severe hypertriglyceridemia including, but not limited to, sustained chylomicronemia.

[0035] The present invention refers to measurement of chylomicrons, vitamin E, and / or triglycerides, etc. in the blood of a subject. In the context of the present invention the term “blood” broadly encompasses whole blood as well as blood components such as plasma and serum. It is within the skills of those in the art to determine whether measurements must be carried out in whole blood, serum and / or plasma.

[0036] As used herein, the term “triglyceride”” or “TG” refers to a tri-ester consisting of a glycerol bound to three fatty acid molecules. As is known, triglycerides are the main constituents of body fat in humans and other vertebrates. Triglycerides are transported by the largest and most lipid-rich of the lipoprotein particles, namely, chylomicrons and very low density lipoproteins (VLDL).

[0037] As used herein, the term “hypertriglyceridemia” refers to a condition in which triglyceride levels are elevated (e.g., 1 ,7 mmoL / L or higher). The term “subject suffering from hypertriglyceridemia” refers to a subject having (e.g., diagnose) or susceptible to having hypertriglyceridemia. The term “subject suffering from chylomicronemia refers to an adult subject having a fastening triglyceride level of more than >10 mmol / L. As is known, a normal fastening triglyceride level in adults is below 1 ,7 mmol / L and, in embodiments, a triglyceride level above 10 mmol / L is considered chylomicrons-related triglycerides.

[0038] As used herein, the term “lipoproteins” refers to particles comprising a core of triglyceride and cholesteryl ester and a coat comprising phospholipid, free cholesterol, and apolipoproteins. There are four major lipoprotein particles: chylomicrons (CMs), very low-density lipoprotein (VLDL), LDL (low-density lipoprotein), and high-density lipoprotein (HDL).

[0039] As used herein, the term “chylomicrons” refers to one of the major lipoprotein particles also known as ultra low-density lipoproteins (ULDL). Typically, chylomicrons are large triglyceride-rich lipoproteins produced in enterocytes to transport dietary lipids. Chylomicrons are composed of triglycerides (about 85-92%), phospholipids (about 6- 12%), cholesterol (about 1-3%), and proteins (about 1-2%). As used herein, the term “chylomicronemia” refers to a condition wherein abnormally elevated levels of triglycerides in the chylomicron pathway are found in the blood (e.g., TG >10 mmol / l). The term “sustained chylomicronemia” refers to a condition wherein the chylomicronemia is persistent and confirmed at least 3 times, 3 months apart.

[0040] As used herein interchangeably, the terms “chylomicrons-related triglycerides” and “chylomicron-TG “ refer to those triglycerides that are transported by chylomicrons, and not transported by the very low density lipoproteins (VLDL). Likewise, the term “triglycerides-related VLDL” refers to those triglycerides that are transported by the VLDL, and not transported by chylomicrons.Assessment of chylomicrons-related triglycerides in blood

[0041] The chylomicron transports exogenous or dietary fat in blood, and in accordance with the present invention, the level of vitamin E may be used as a clinical marker of the blood level of chylomicrons-related triglycerides.

[0042] Accordingly, one aspect of the present invention concerns a method to assess a blood level of chylomicrons-related triglycerides in a subject suffering from severe hypertriglyceridemia. In one embodiment, the method comprises measuring a level of vitamin E in the subject’s blood, and in accordance with the present invention, the level of vitamin E is indicative of the blood level of chylomicrons-related triglycerides.

[0043] In embodiments, the subject suffers from severe hypertriglyceridemia (e.g., TG >5 mmol / l). In embodiments, the subject suffers from sustained chylomicronemia (TG >10 mmol / l).Triglycerides issued from the exogenous pathway or from the endogenous pathway

[0044] One aspect of the present invention concerns using vitamin E to distinguish between the two pathways associated with blood triglycerides, i.e., triglycerides and lipoproteins issued from the exogenous pathway versus triglycerides issued from the endogenous pathway.

[0045] Particularly, one aspect of the present invention concerns a method to distinguish between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway, preferably in blood of a subject suffering from hypertriglyceridemia (e.g., a subject displaying a fastening triglyceride level of more than about 5 mmol / L or more than about 10 mmol / L). In one embodiment the method comprises measuring a level of vitamin E in the subject’s blood, and the level of vitamin E is indicative of levels of triglycerides and lipoproteins issued from the exogenous pathway.

[0046] In embodiments, the blood level of triglycerides and lipoproteins issued from the exogenous pathway is directly correlated to the measured level of vitamin E. In embodiments, measuring the level of vitamin E in the subject’s blood provides for and / or confirms a diagnostic of chylomicronemia.

[0047] In embodiments, the method further comprises administering vitamin E to the subject and measuring thereafter the blood concentration of vitamin E. In embodiments, the method comprises administering about 2 500 Ul to about 10 000 Ul of vitamin E, or about 5 000 Ul of vitamin. The blood concentration of vitamin E may be measured at any suitable time, for instance about 1 to 12 hours, or 1 hour, or 2 hours, or about 4 hours, or about 6 hours, or about 12 hours, or about 24 hours after the administration of vitamin E. The vitamin E may be administered by any suitable route (e.g., orally) and, in embodiments, the dosage may vary according to age. The vitamin E may be administered in a single dose, or it may be administered in multiple smaller doses extending over a longer period. For example, in patients with abetalipoproteinemia, the prescribed dosage is 9600UI, using 400UI capsids.Efficacy of hypolipidemic treatments

[0048] Another aspect of the present invention concerns using vitamin E for assessing whether hypolipidemic treatments are effective or not, and to which extent. The invention also provides for adjusting a treatment plan based on vitamin E measurements and / or desired TG level. As is known, in accordance with medical consensus, an optimal level is < 1.2 mmol / L (about 100 mg / dL), a borderline level is 1.2 -1.7 mmol / L (100-150 mg / dL), a moderately elevated is 1.7 - 5.7 mmol / L (150-500 mg / dL), a severe level is 5.7-10 mmol / L (500-880 mg / dL) and an extreme level is >10 mmol / L (>880 mg / dL) (Ginsberg H.N, et al., European Heart Journal (2021) 42, 4791-4806).

[0049] Particularly, one aspect of the invention concerns a method to assess efficacy of an hypolipidemic therapy, preferably in a subject suffering from severe hypertriglyceridemia. In one embodiment the method comprises: (i) measuring a first level of vitamin E in the subject’s blood prior to beginning of the hypolipidemic therapy; and (ii) measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy.

[0050] In embodiments, the subject displays a fastening triglyceride level of more than about 5 mmol / l prior to the beginning of the therapy. In embodiments, the subject displays a fastening triglyceride level of more than about 10 mmol / L prior to the beginning of the therapy.

[0051] In embodiments, the method further comprises monitoring levels of triglycerides and vitamin E during the hypolipidemic therapy. In embodiments, the second level of vitamin E is measured about 1 week, or about 2 weeks, or about 1 month, or about 2 months, after beginning of the hypolipidemic therapy. In embodiments, levels of vitamin E and / or triglycerides are measured continuously during the therapy (e.g., daily, weekly, biweekly, monthly, bimonthly, a few times per year, etc.) and the treatment plan is adjusted accordingly.

[0052] In embodiments, measuring level of vitamin E in the subject further comprises a preliminary step of administering vitamin E prior to each of the measurement. In one embodiment, vitamin E is administered prior to beginning of the hypolipidemic therapy andafter beginning of the hypolipidemic therapy. In embodiments, a dose of vitamin E to be administered is about 2 500 Ul to about 10 000 Ul of vitamin E, or about 5 000 Ul of vitamin E.

[0053] In embodiments, the hypolipidemic therapy comprises a very low-fat diet. In embodiments, the hypolipidemic therapy comprises administering a suitable hypolipidemic drug. Existing hypolipidemic drugs include those compounds from the following classes of compounds: fibrates, APOC3 inhibitors, ANGPLT3 inhibitors, omega-3 agents. Particular examples existing products or compounds currently in use or development for the treatment of hypolipidemia include, but is not limited to, fish oils, medium chain triglycerides (MCT) oil, omega 3, clofibrate, gemfibrozil, fenofibrate, niacin, APOC3 antisense oligonucleotides (ASO), ANGPTL3 ASO, anti-ANGPLT3 antibodies, etc.

[0054] In embodiments, the method allows to distinguish between (i) an effect of the hypolipemic therapy on triglycerides and lipoproteins issued from the exogenous pathway versus (ii) an effect of the hypolipemic therapy on triglycerides issued from the endogenous pathway.Risks of acute pancreatitis

[0055] As is known, chylomicronemia increases the risks of acute pancreatitis. Accordingly, another aspect of the present invention concerns using vitamin E levels for assessing the risks of acute pancreatitis in subjects. Particularly, one aspect of the invention concerns a method to assess a risk of acute pancreatitis in a subject suffering from hypertriglyceridemia. In one embodiment the method comprises measuring a level of vitamin E in the subject’s blood, and in accordance with this method, the level of vitamin E is indicative of the blood level of chylomicrons, and a higher level of chylomicrons is indicative of a higher risk of acute pancreatitis for the subject.

[0056] In accordance with the present invention normal vitamin E levels are defined in Table 1 below:Table 1: Lower limit normal (LLN) and Upper limit normal (ULN) of vitamin E levels in humans*These values may vary from one country to another

[0057] In embodiments, an elevated level of vitamin E which is twice (2x) the upper limit normal (ULN) for the age is indicative of a higher risk of acute pancreatitis for the subject.

[0058] In embodiments, the subject displays a fastening triglyceride level of more than about 5 mmol / l. In embodiments, the subject displays a fastening triglyceride level of more than about 10 mmol / L.

[0059] Depending on the risk assessment, the medical practitioner can take a proper course of action. For instance, MCT oil supplement, severe low-fat diet or agents decreasing TG.Vitamin E deficiency

[0060] Vitamin E deficiency is a rare condition which can cause nerve problems. The deficiency is usually due to an underlying problem with digesting dietary fat rather than from a diet abnormally low in vitamin E.

[0061] Another aspect of the present invention concerns using chylomicrons for assessing an amount of vitamin E that is bioavailable and therefore, for assessing a need of vitamin E in subjects. Particularly, one aspect of the invention concerns a method toassess a need of vitamin E in a subject suffering from hypertriglyceridemia. In one embodiment the subject suffers from severe hypertriglyceridemia and method comprises measuring a level of chylomicrons in the blood of the subject and, in accordance with the present method, the level of chylomicrons is indicative of a level of vitamin E that is not bioavailable to the subject, and the level of vitamin E that is not bioavailable is indicative of a need in vitamin E.

[0062] In one embodiment, the subject suffers from abetalipoproteinemia and thus requires a supplement of vitamin E at least because in that subject the vitamin E is not bioavailable since it is contained in the chylomicrons.

[0063] In embodiments, the method further comprises administering vitamin E to a subject displaying an abnormally low level of vitamin E. In embodiments the subject requiring vitamin E may display a fastening triglyceride level of more than about 5 mmol / l (e.g., more than about 10 mmol / l, or more than about 20 mmol / l) and a level of vitamin E that is below expected normal values (see Table 1 above). In embodiments, the subject requiring vitamin E displays a fastening triglyceride level of more than about 10 mmol / l or more than about 20 mmol / l and a level of vitamin E that is 2 x below LLN values shown in Table 1.

[0064] The vitamin E may be administered by any suitable route (e.g., orally). In embodiments the amounts of vitamin E that is administered to the subject may be of about 5 000 Ul to about 20 000 Ul per day, or about 7 500 Ul to about 15 000 Ul per day, or about 10 000 Ul per day. The vitamin E may be administered in a single dose, or it may be administered in multiple smaller doses extending over a longer period (weeks, months, years). In one particular embodiment, the subject is administered 24 doses per day, each dose comprising about 400 Ul of vitamin E.Kits comprising vitamin E

[0065] Another aspect of the invention concerns kits comprising vitamin E.

[0066] In one embodiment, the kit comprises multiple doses of vitamin E, such kit including for instance blister packages, capsule strips, reclosable bottles, powder packs,and the like. These may comprise a certain amount of vitamin E provided in a powder form (e.g., 500 mg, 1 000 mg, 2500 mg, 5 000 mg, 1000 mg, 20 000 mg or more of vitamin E), in a liquid form (e.g., 50 ml, 100 ml, 500 ml, 1 000 ml or more of an edible solution comprising vitamin E) or a number of vitamin capsules or tablets (e.g., 10, 25, 50 or more of such capsules or tablets) each comprising a certain dose of vitamin E (e.g., about 100 Ul to about 1 000 Ul of vitamin E, about 250 Ul to about 500 Ul of vitamin E, or about 400 Ul of vitamin E). Such kit can also advantageously include instructions in the form of a pamphlet or any other printed support, these instructions indicating the quantities of vitamin E to be administered, the dosage regimen, information about the route of administration, how to mix the components (e.g., if used in a powder form or a liquid concentrate), etc.

[0067] In one particular embodiment, the kit is configured for assessing efficacy of an hypolipidemic therapy in a subject suffering from severe hypertriglyceridemia. Accordingly, the kit is configured with the objective of measuring a first level of vitamin E in the subject’s blood prior to beginning of the hypolipidemic therapy, and then measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy.

[0068] In accordance with that particular embodiment, the kit comprises (i) a first strip tab comprising one or more capsules or tablets of vitamin E to be taken prior to the beginning of the hypolipidemic therapy and (ii) a second strip tab comprising one or more capsules or tablets of vitamin E to be taken after beginning of the hypolipidemic therapy. In embodiments each strip comprises about 2 to 24 capsules or tablets, each of them containing about 400 Ul of vitamin E. In embodiments the kit also comprises a package for the strips and written instructions.

[0069] Kits in accordance with the present invention may also find additional applications including, but not limited to, in the course of reducing to practice the methods described hereinbefore.EXAMPLESExample 1 : Pediatric case

[0070] Babies less than one year old with a sustained severe hypertriglyceridemia may present a LPL deficiency (LPLD) and they accumulate chylomicrons since they cannot hydrolyze the TGs contained in the chylomicrons.

[0071] Since vitamin E is carried by chylomicrons, in accordance with the present invention, measurement of vitamin E in babies, infants and pediatric patients can confirm that the hypertriglyceridemia depends on the accumulation of chylomicrons when vitamin E values are elevated. In accordance with the present invention, vitamin E measurement is also a clinical marker of the effectiveness of hypolipidemic treatment on chylomicron- triglyceride specifically by assessing a decrease / fluctuation in vitamin E levels.

[0072] This is illustrated by the following case of a baby for which a severe hypertriglyceridemia was diagnosed at the age of 2 months. As shown in Table 2, the blood (plasma) vitamin E levels at 2 months were much higher than the normal vitamin E values for a healthy 2 to 6-month-old baby (a measured value of 121 mol / L instead of a normal value of 5-14 pmol / L). However, changing the baby’s diet by providing a LPLD- adapted milk formula resulted in a significant decrease in both, vitamin E levels and triglycerides levels.Table 2: Correlation of plasma TG levels vs vitamin E in a baby

[0073] These results confirms that plasma Vitamin E levels are elevated in presence of chylomicronemia and that Vitamin E levels fluctuate according to TG levels.Example 2: Adult cases

[0074] In adults, normal values of vitamin E in the blood are higher (13-43 pmol / L) than in children. Sustained severe hypertriglyceridemia in adults may depend on TG in the lipoproteins of the exogenous pathway (chylomicron pathway) or the endogenous pathway (VLDL pathway and remnants).

[0075] According to the present invention, measurements of vitamin E in the blood of adults allow an evaluation of the contribution of TG in exogenous lipoproteins (chylomicrons) vs the endogenous pathway (VLDL and remnants). The present invention also provides for evaluation of the effectiveness of a therapeutic treatment. Table 3 provides a demonstration of these concepts in hypertriglyceridemic adult subjects.Table 3: Vitamin E vs TG in adults with hypertriglyceridemia* Chylomicrons (CM) exogenous pathway vs very low density lipoproteins (VLDL). endogenous pathway

[0076] As shown in Table 3, subjects 5, 6, 7 illustrate that when vitamin E levels are normal, TG levels represent a significant proportion of endogenous (VLDL) - related TGs. This has been confirmed, at least for subject 5, by measuring APO-B 100 levels and via lipoproteins ultracentrifugation.

[0077] Headings are included herein for reference and to aid in locating certain sections. These headings are not intended to limit the scope of the concepts described therein, and these concepts may have applicability in other sections throughout the entire specification. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

[0078] The singular forms “a”, “an” and “the” include corresponding plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a subject" includes one or more of such subjects and reference to "the method" includes reference to equivalent steps and methods known to those of ordinary skill in the art that could be modified or substituted for the methods described herein.

[0079] Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, concentrations, properties, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present specification and attached claims are approximations that may vary depending upon the properties sought to be obtained. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the embodiments are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors resulting from variations in experiments, testing measurements, statistical analyses and such.

[0080] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the present invention and scope of the appended claims.

Claims

CLAIMS:1 . A method to assess a blood level of chylomicrons-related triglycerides in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood, wherein the level of vitamin E is indicative of the blood level of chylomicrons- related triglycerides.

2. The method of claim 1 , wherein the blood level of chylomicrons-related triglycerides is directly correlated to the measured level of vitamin E.

3. The method of claim 1 or 2, wherein the subject comprises triglycerides blood levels of >5 mmol / l.

4. The method of claim 1 , wherein the subject comprises triglycerides blood levels of >10 mmol / l.

5. A method to distinguish between (i) triglycerides and lipoproteins issued from the exogenous pathway versus (ii) triglycerides issued from the endogenous pathway in blood of a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood, wherein the level of vitamin E is indicative of levels of triglycerides and lipoproteins issued from the exogenous pathway.

6. The method of claim 5, wherein the blood level of triglycerides and lipoproteins issued from the exogenous pathway is directly correlated to the measured level of vitamin E.

7. The method of claim 5 or 6, wherein measuring the level of vitamin E in the subject’s blood provides for and / or confirms a diagnostic of chylomicronemia.

8. The method of any one of claims 5 to 7, wherein the subject suffering from hypertriglyceridemia displays a fastening triglyceride level of more than about 5 mmol / l.

9. The method of any one of claims 5 to 8, wherein the subject suffering from hypertriglyceridemia displays a fastening triglyceride level of more than about 10 mmol / L.

10. The method of any one of claims 5 to 9, further comprising administering vitamin E to the subject and measuring thereafter the blood concentration of vitamin E.11 . The method of claim 10, wherein the vitamin E is administered orally.

12. The method of claim 10, comprising administering about 2 500 Ul to about 10 000 Ul of vitamin E.

13. The method of claim 10, wherein the blood concentration of vitamin E is measured about 1 hour to about 12 hours after said administration of vitamin E.

14. A method to assess efficacy of an hypolipidemic therapy in a subject suffering from severe hypertriglyceridemia, the method comprising:(i) measuring a first level of vitamin E in the subject’s blood prior to beginning of the hypolipidemic therapy;(ii) measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy.

15. The method of claim 14, wherein the subject displays a fastening triglyceride level of more than about 5 mmol / l prior to the beginning of the therapy.

16. The method of claim 14, wherein the subject displays a fastening triglyceride level of more than about 10 mmol / L prior to the beginning of the therapy.

17. The method of any one of claims 14 to 16, further comprising monitoring levels of triglycerides during the hypolipidemic therapy.

18. The method of any one of claims 14 to 16, further comprising adjusting a treatment plan of the hypolipidemic therapy, wherein said adjusting takes into consideration said vitamin E measurements.

19. The method of any one of claims 14 to 18, wherein the second level of vitamin E is measured about 1 week, or about 2 weeks, or about 1 month, or about 2 months, after beginning of the hypolipidemic therapy.

20. The method of any one of claims 14 to 19, wherein the levels of vitamin E and / or levels of triglycerides are measured continuously during the therapy.21 . The method of any one of claims 14 to 20, further comprising administering to the subject vitamin E prior to beginning of the hypolipidemic therapy and after beginning of the hypolipidemic therapy.

22. The method of any one of claims 14 to 20, further comprising administering to the subject vitamin E prior to each measurement of vitamin E.

23. The method of claim 21 or 22, comprising administering about 2 500 Ul to about 10 000 Ul of vitamin E.

24. The method of any one of claims 14 to 23, wherein the hypolipidemic therapy comprises providing a very low-fat diet and / or administering an hypolipidemic drug or hypolipidemic compound.

25. The method of claim 24, wherein the hypolipidemic drug or hypolipidemic compound is selected from the group consisting of fish oils, omega 3, medium chain triglycerides (MCT) oil, clofibrate, gemfibrozil, fenofibrate, niacin, APOC3 antisense oligonucleotides (ASO), ANGPTL3 ASO, and anti-ANGPLT3 antibodies.

26. The method of any one of claims 14 to 25, wherein said method allows to distinguish between (i) an effect of the hypolipemic therapy on triglycerides and lipoproteins issued from the exogenous pathway versus (ii) an effect of the hypolipemic therapy on triglycerides issued from the endogenous pathway.

27. A method to assess a risk of acute pancreatitis in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of vitamin E in the subject’s blood; wherein said level of vitamin E is indicative of the blood level of chylomicrons; andwherein a higher level of chylomicrons is indicative of a higher risk of acute pancreatitis for the subject.

28. The method of claim 27, wherein a level of vitamin E of 2 times the upper limit normal (ULN) is indicative of a higher risk of acute pancreatitis for the subject.

29. The method of claim 27 or 28, wherein the subject displays a fastening triglyceride level of more than about 5 mmol / l.

30. The method of claim 27 or 28, wherein the subject displays a fastening triglyceride level of more than about 10 mmol / L.

31. A method to assess a need of vitamin E in a subject suffering from severe hypertriglyceridemia, the method comprising measuring a level of chylomicrons in the blood of the subject; wherein said level of chylomicrons is indicative of a level of vitamin E that is not bioavailable to the subject; and wherein the level of vitamin E that is not bioavailable is indicative of a need of vitamin E.

32. The method of claim 31 , wherein said subject suffers from abetalipoproteinemia and requires vitamin E.

33. The method of claim 31 , wherein the subject displays a fastening triglyceride level of more than about 10 mmol / l and a level of vitamin E that is below normal values.

34. The method of claim 31 , wherein the subject displays a fastening triglyceride level of more than about 20 mmol / l and a level of vitamin E that is 2 times below lower limit normal (LLN).

35. The method of any one of claims 31 to 34, further comprising administering vitamin E to a subject displaying an abnormally low level of vitamin E.

36. The method of claim 35, comprising administering about 5 000 Ul to about 20 000 Ul of vitamin E per day.

37. The method of claim 35 or 36, wherein the vitamin E is administered orally.

38. A kit for assessing efficacy of an hypolipidemic therapy in a subject suffering from severe hypertriglyceridemia, the kit comprising multiple doses of vitamin E, and wherein said doses of vitamin E are comprised in a blister package, a capsule strip, a reclosable bottle, a powder pack.

39. The kit of claim 38, wherein the kit comprises: (i) a first strip tab comprising one or more capsules or tablets of vitamin E to be taken prior to the beginning of the hypolipidemic therapy; and (ii) a second strip tab comprising one or more capsules or tablets of vitamin E to be taken after beginning of the hypolipidemic therapy.

40. The kit of claim 38 or 39, comprising at least 2 capsules, tablets or powder pack of vitamin E.41 . The kit of any one of claims 38 to 40, wherein each strip tab comprises at least 12 capsules or tablets of vitamin E.

42. The kit of any one of claims 38 to 41 , wherein each of the capsule, tablet or powder pack comprises about 100 Ul to about 1 000 Ul of vitamin E.

43. The kit of any one of claims 38 to 42, wherein each of the capsule, tablet or powder pack comprises about 400 Ul of vitamin E.

44. Use of a vitamin E blood measurement as a clinical marker of a subject’s blood level of chylomicrons-related triglycerides.

45. Use of a vitamin E blood measurement to clinically distinguish in a subject between(i) triglycerides and lipoproteins issued from the exogenous pathway versus(ii) triglycerides issued from the endogenous pathway.

46. Use of a vitamin E blood measurement in a subject suffering from severe hypertriglyceridemia for assessing efficacy of hypolipidemic therapies.

47. Use of a vitamin E blood measurement in a subject suffering from severe hypertriglyceridemia to assess the subject’s risks of an acute pancreatitis.

48. Use of a chylomicrons blood measurement in a subject suffering from severe hypertriglyceridemia to assess the subject’s need in vitamin E.

49. A method for treating a subject suffering from severe hypertriglyceridemia, the method comprising:(i) measuring a first level of vitamin E in the subject’s blood prior to beginning of an hypolipidemic therapy;(ii) measuring a second level of vitamin E in the subject’s blood after beginning of the hypolipidemic therapy; wherein said measurements of vitamin E provide a clinical marker of the subject’s blood level of chylomicrons-related triglycerides; and(iii) providing a treatment plan taking into consideration the subject’s blood level of chylomicrons-related triglycerides.

50. The method of claim 49, wherein said treatment plan comprises treating said subject for hypertriglyceridemia.

51. The method of claim 49, wherein said treatment plan comprises treating said subject for chylomicronemia.

52. The method of claim 49, wherein said treatment plan comprises treating said subject for acute pancreatitis or other morbidities.

53. The method of claim 49, wherein said treatment plan comprises treating said subject for vitamin E deficiency.

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