Familial Familial HypercholesterolemiaHypercholesterolemia

Honors GeneticsHonors Genetics

20062006

Gel InterpretationGel Interpretation

Measure the distance from Measure the distance from the well to the bands( DNA the well to the bands( DNA fragments in the marker)fragments in the marker)

Graph using the marker sizes Graph using the marker sizes for Lambda Hind IIIfor Lambda Hind III

Interpolate the sizes of the Interpolate the sizes of the bands for the patients, bands for the patients, Normal and FHC control to Normal and FHC control to verify your visual verify your visual observations and impressionsobservations and impressions

Hypercholesterolemia Hypercholesterolemia Gel ResultsGel Results

N

FHC

1

2

3

marker

Gel InterpreationGel Interpreation

Patient 1 – Homozygous Patient 1 – Homozygous recessive – both copies of the recessive – both copies of the normal LDLR receptornormal LDLR receptor

Patient 2 – Heterozygous for Patient 2 – Heterozygous for the LDLR receeptorthe LDLR receeptor

Patient 3- Homozygous Patient 3- Homozygous dominant for the LDLR dominant for the LDLR receptor – The most severe receptor – The most severe form of the diseaseform of the disease

What is cholesterol??What is cholesterol?? Cholesterol is one of the body's fats Cholesterol is one of the body's fats

(lipids). Cholesterol and another (lipids). Cholesterol and another lipid, triglyceride, are important lipid, triglyceride, are important building blocks in the structure of building blocks in the structure of cells and are also used in making cells and are also used in making hormones and producing energy. hormones and producing energy.

To some extent, the cholesterol To some extent, the cholesterol level in blood depends on what you level in blood depends on what you eat but it is mainly dependent on eat but it is mainly dependent on how the body makes cholesterol in how the body makes cholesterol in the liver.the liver.

Cholesterol and the Cholesterol and the Steroid RingsSteroid Rings

Desirable cholesterol Desirable cholesterol levelslevels

DesirableDesirable — Less than 200 mg/dL — Less than 200 mg/dLBorderline high riskBorderline high risk — 200–239  — 200–239 mg/dLmg/dLHigh riskHigh risk — 240 mg/dL and over  — 240 mg/dL and over

If your total cholesterol is less than If your total cholesterol is less than 200 mg/dL, your heart attack risk is 200 mg/dL, your heart attack risk is relatively low, unless you have other relatively low, unless you have other risk factors. Even with a low risk, it's risk factors. Even with a low risk, it's still smart to eat foods low in still smart to eat foods low in saturated fat and cholesterol, and saturated fat and cholesterol, and also get plenty of physical activity. also get plenty of physical activity. Have your cholesterol levels Have your cholesterol levels measured every five years — or more measured every five years — or more often if you're a man over 45 or a often if you're a man over 45 or a woman over 55 woman over 55

Low density Low density lipoproteinslipoproteins

This is sometimes called “ bad “ This is sometimes called “ bad “ cholesterolcholesterol

the blood, and the cholesterol from the blood, and the cholesterol from LDL is the main source of damaging LDL is the main source of damaging buildup and blockage in the arteries. buildup and blockage in the arteries.

Thus, the more LDL-cholesterol you Thus, the more LDL-cholesterol you have in your blood, the greater your have in your blood, the greater your risk of heart disease. If you have risk of heart disease. If you have heart disease or are at high risk for heart disease or are at high risk for developing it and your LDL is 100 developing it and your LDL is 100 mg/dLor higher, your cholesterol mg/dLor higher, your cholesterol may well be too high for you. may well be too high for you.

High Density High Density LipoproteinsLipoproteins

HDL carry cholesterol in the blood from HDL carry cholesterol in the blood from other parts of the body back to the other parts of the body back to the liver, which leads to its removal from liver, which leads to its removal from the body. the body.

So HDL help keep cholesterol from So HDL help keep cholesterol from building up in the walls of the arteries. building up in the walls of the arteries. If your level of HDL-cholesterol is below If your level of HDL-cholesterol is below 40 mg/dL, you are at substantially 40 mg/dL, you are at substantially higher risk for heart disease. higher risk for heart disease.

The higher your HDL-cholesterol, the The higher your HDL-cholesterol, the better. The average HDL-cholesterol for better. The average HDL-cholesterol for men is about 45 mg/dL, and for women men is about 45 mg/dL, and for women it is about 55 mg/dL.it is about 55 mg/dL.

Tests in the Lipid Tests in the Lipid ProfileProfile

Total Cholesterol Total Cholesterol HDL Cholesterol HDL Cholesterol LDL Cholesterol LDL Cholesterol VLDL Cholesterol VLDL Cholesterol Triglycerides Triglycerides Apolipoprotein A1Extended Lipid Apolipoprotein A1Extended Lipid

Profile. Profile. Apolipoprotein B Apolipoprotein B Lipoprotein(a) Lipoprotein(a)

Before withdrawing blood for Before withdrawing blood for lipid profile :-lipid profile :-

          a. 12 - 14 hours fasting a. 12 - 14 hours fasting sample is required.sample is required.

        b. Individuals should have b. Individuals should have been on their usual diet.been on their usual diet.

Case HistoryCase Historyreferene - OMIMreferene - OMIM

European populations. European populations. Like the French Canadians, the South Afrikaners Like the French Canadians, the South Afrikaners

appear to have a unique form of mutation in the appear to have a unique form of mutation in the LDLR gene consistent with founder effect (Brink LDLR gene consistent with founder effect (Brink et al., 1987). Because of the presumed role of et al., 1987). Because of the presumed role of founder effect on the high frequency of familial founder effect on the high frequency of familial hypercholesterolemia in South Africa, it is not hypercholesterolemia in South Africa, it is not surprising that Kotze et al. (1987) found a surprising that Kotze et al. (1987) found a predominance of 2 haplotypes in 27 informative predominance of 2 haplotypes in 27 informative families with FH. In a study of homozygotes families with FH. In a study of homozygotes from the Afrikaner population in South Africa, from the Afrikaner population in South Africa, Leitersdorf et al. (1989) found that 2 mutations Leitersdorf et al. (1989) found that 2 mutations account for more than 95% of the mutant LDL account for more than 95% of the mutant LDL receptor genes. Both mutations were basepair receptor genes. Both mutations were basepair substitutions that resulted in a single amino acid substitutions that resulted in a single amino acid change and both could be detected readily with change and both could be detected readily with PCR and restriction analysis. The findings were PCR and restriction analysis. The findings were considered consistent with the high frequency considered consistent with the high frequency of FH being due to founder effect. of FH being due to founder effect.

Chromosome – 19.13.2Chromosome – 19.13.2

Start:Start:11,061,13211,061,132 bp from bp from pterEnd:pterEnd:11,105,49011,105,490 bp from bp from pterSize:pterSize:44,35844,358 bases bases

Gene ExpressionGene Expression

SNP – Single nucleotide SNP – Single nucleotide polymorphismpolymorphism

CTATGACACCCTATGACACCA/GA/GTCATCAGCAGTCATCAGCAG rs11669576 rs11669576 F11083300(+)CCACACGAAGF11083300(+)CCACACGAAGA/GA/GCCTGCAAGGC CCTGCAAGGC rs5928 rs5928

F11101240(+)AAGAACTGGCF11101240(+)AAGAACTGGCA/GA/GGCTTAAGAAC 1R/Q ns 2A:0.01MN 310 --GCTTAAGAAC 1R/Q ns 2A:0.01MN 310 -- rs5930 rs5930 F,C,A11085265(+)TCATCAGCAGF,C,A11085265(+)TCATCAGCAGA/GA/GGACATCCAGG 1R/R syn 3A:0.37MN GACATCCAGG 1R/R syn 3A:0.37MN

EA 1810 --EA 1810 -- rs5927 rs5927 F,C,A11094941(+)CAACCACCCGF,C,A11094941(+)CAACCACCCGA/GA/GCCTGTTCCCG 1R/R syn 2A:0.08MN CCTGTTCCCG 1R/R syn 2A:0.08MN

EA 1620 EA 1620 rs5925 rs5925 F,C,A11091881(+)AGGATATGGTF,C,A11091881(+)AGGATATGGTC/TC/TCTCTTCCACA 1V/V syn 3C:0.18MN EA CTCTTCCACA 1V/V syn 3C:0.18MN EA

1810 --1810 -- rs688 rs688 F,C,A11088602(+)TCGATGTCAAF,C,A11088602(+)TCGATGTCAAC/TC/TGGGGGCAACC 1N/N syn 1T:0.32MN GGGGGCAACC 1N/N syn 1T:0.32MN

126 126 rs5929 rs5929 F,C11087800(+)GGGGAACTCCF,C11087800(+)GGGGAACTCCC/TC/TGCCAAGATCA 1P/P syn 2T:0.04MN 310 GCCAAGATCA 1P/P syn 2T:0.04MN 310

rs1799898 rs1799898 F,C11088554(+)GTGGCCGCCTF,C11088554(+)GTGGCCGCCTC/TC/TTACTGGGTTG 1L/L syn 0--------TACTGGGTTG 1L/L syn 0-------- rs5933 rs5933 F,C11085090(+)GCAATAGAATF,C11085090(+)GCAATAGAATC/TC/TTACTGGTCTG TACTGGTCTG

HGMDHGMD

http://archive.uwcm.ac.uk/uwhttp://archive.uwcm.ac.uk/uwcm/mg/search/119362.htmlcm/mg/search/119362.html

This data base shows all the This data base shows all the different mutationsdifferent mutations

For Familial For Familial hypercholesterolemia there hypercholesterolemia there are 806 mutationsare 806 mutations

457 mutations are missense 457 mutations are missense and nonsenseand nonsense

Substitution mutationsSubstitution mutations

GGG-AGG Gly-Arg GGG-AGG Gly-Arg HypercholesterolaemiaHypercholesterolaemia

GCG-GAG Ala-Glu GCG-GAG Ala-Glu HypercholesterolaemiaHypercholesterolaemia

CTC-CCC Leu-Pro CTC-CCC Leu-Pro HypercholesterolaemiaHypercholesterolaemia

cGAG-TAG Glu-Term cGAG-TAG Glu-Term HypercholesterolaemiaHypercholesterolaemia

RCSB – The LDLR RCSB – The LDLR receptorreceptor

Sequence for LDLR Sequence for LDLR proteinprotein

1100 2 200 3 300 4 400 5 500 6 600 MGPWGWKLRW TVALLLAAAG TAVGDRCERN MGPWGWKLRW TVALLLAAAG TAVGDRCERN EFQCQDGKCI SYKWVCDGSA ECQDGSDESQ 7EFQCQDGKCI SYKWVCDGSA ECQDGSDESQ 700 8 800 9 900 10 1000 11 1100 12 1200 ETCLSVTCKS GDFSCGGRVN RCIPQFWRCD GQVDCDNGSD ETCLSVTCKS GDFSCGGRVN RCIPQFWRCD GQVDCDNGSD EQGCPPKTCS QDEFRCHDGK 13EQGCPPKTCS QDEFRCHDGK 1300 14 1400 15 1500 16 1600 17 1700 18 1800 CISRQFVCDS CISRQFVCDS DRDCLDGSDE ASCPVLTCGP ASFQCNSSTC IPQLWACDND DRDCLDGSDE ASCPVLTCGP ASFQCNSSTC IPQLWACDND PDCEDGSDEW 19PDCEDGSDEW 1900 20 2000 21 2100 22 2200 23 2300 24 2400 PQRCRGLYVF PQRCRGLYVF QGDSSPCSAF EFHCLSGECI HSSWRCDGGP DCKDKSDEEN QGDSSPCSAF EFHCLSGECI HSSWRCDGGP DCKDKSDEEN CAVATCRPDE 25CAVATCRPDE 2500 26 2600 27 2700 28 2800 29 2900 30 3000 FQCSDGNCIH FQCSDGNCIH GSRQCDREYD CKDMSDEVGC VNVTLCEGPN KFKCHSGECI GSRQCDREYD CKDMSDEVGC VNVTLCEGPN KFKCHSGECI TLDKVCNMAR 31TLDKVCNMAR 3100 32 3200 33 3300 34 3400 35 3500 36 3600 DCRDWSDEPI DCRDWSDEPI KECGTNECLD NNGGCSHVCN DLKIGYECLC PDGFQLVAQR KECGTNECLD NNGGCSHVCN DLKIGYECLC PDGFQLVAQR RCEDIDECQD 37RCEDIDECQD 3700 38 3800 39 3900 40 4000 41 4100 42 4200 PDTCSQLCVN PDTCSQLCVN LEGGYKCQCE EGFQLDPHTK ACKAVGSIAY LFFTNRHEVR LEGGYKCQCE EGFQLDPHTK ACKAVGSIAY LFFTNRHEVR KMTLDRSEYT 43KMTLDRSEYT 4300 44 4400 45 4500 46 4600 47 4700 48 4800 SLIPNLRNVV ALDTEVASNR SLIPNLRNVV ALDTEVASNR IYWSDLSQRM ICSTQLDRAH GVSSYDTVIS RDIQAPDGLA 49IYWSDLSQRM ICSTQLDRAH GVSSYDTVIS RDIQAPDGLA 4900 50 5000 51 5100 525200 53 5300 54 5400 VDWIHSNIYW TDSVLGTVSV ADTKGVKRKT VDWIHSNIYW TDSVLGTVSV ADTKGVKRKT LFRENGSKPR AIVVDPVHGF MYWTDWGTPA 55LFRENGSKPR AIVVDPVHGF MYWTDWGTPA 5500 56 5600 57 5700 58 5800 59 5900 606000 KIKKGGLNGV DIYSLVTENI QWPNGITLDL LSGRLYWVDS KIKKGGLNGV DIYSLVTENI QWPNGITLDL LSGRLYWVDS KLHSISSIDV NGGNRKTILE 61KLHSISSIDV NGGNRKTILE 6100 62 6200 63 6300 64 6400 65 6500 66 6600 DEKRLAHPFS DEKRLAHPFS LAVFEDKVFW TDIINEAIFS ANRLTGSDVN LLAENLLSPE DMVLFHNLTQ LAVFEDKVFW TDIINEAIFS ANRLTGSDVN LLAENLLSPE DMVLFHNLTQ 676700 68 6800 69 6900 70 7000 71 7100 72 7200 PRGVNWCERT TLSNGGCQYL CLPAPQINPH PRGVNWCERT TLSNGGCQYL CLPAPQINPH SPKFTCACPD GMLLARDMRS CLTEAEAAVA 73SPKFTCACPD GMLLARDMRS CLTEAEAAVA 7300 74 7400 75 7500 76 7600 77 7700 78 7800 TQETSTVRLK VSSTAVRTQH TTTRPVPDTS RLPGATPGLT TVEIVTMSHQ TQETSTVRLK VSSTAVRTQH TTTRPVPDTS RLPGATPGLT TVEIVTMSHQ ALGDVAGRGN 79ALGDVAGRGN 7900 80 8000 81 8100 82 8200 83 8300 84 8400 EKKPSSVRAL SIVLPIVLLV EKKPSSVRAL SIVLPIVLLV FLCLGVFLLW KNWRLKNINS INFDNPVYQK TTEDEVHICH 85FLCLGVFLLW KNWRLKNINS INFDNPVYQK TTEDEVHICH 8500 86 8600 NQDGYSYPSR QMVSLEDDVA NQDGYSYPSR QMVSLEDDVA

ReferencesReferences sources: sources: Genetic association of low density lipoprotein recGenetic association of low density lipoprotein rec

eptor and Alzheimer's disease. eptor and Alzheimer's disease.   (2005)(2005)

The modular adaptor protein ARH is required for The modular adaptor protein ARH is required for low density lipoprotein (low density lipoprotein (LDL)bindingLDL)binding and internalization but not for LDL receptor clus and internalization but not for LDL receptor clustering in coated pits. tering in coated pits.   (2004)(2004)

Molecular characterization of familial hypercholeMolecular characterization of familial hypercholesterolemia in Spain: sterolemia in Spain: identificationofidentificationof 39 novel and 77 recurrent mutations in LDLR. 39 novel and 77 recurrent mutations in LDLR.   (2004)(2004)

Harmful effects of increased LDLR expression in Harmful effects of increased LDLR expression in micewithmicewith human APOE*4 human APOE*4 butnotbutnot APOE*3. APOE*3.   (2004)(2004)

Estrogen receptor-alpha and Sp1 interact in the iEstrogen receptor-alpha and Sp1 interact in the induction of the low nduction of the low densitylipoproteindensitylipoprotein-receptor. -receptor.   (2003)(2003)

Generation and initial analysis of more than Generation and initial analysis of more than 15,000 full-length human and mouse cDNA15,000 full-length human and mouse cDNA

ReferencesReferences Comparison of apolipoprotein B Comparison of apolipoprotein B

metabolism in familial defective metabolism in familial defective apolipoproteinB and heterogeneous apolipoproteinB and heterogeneous familial hypercholesterolemia.  familial hypercholesterolemia.  (2002)(2002)

Molecular basis of familial Molecular basis of familial hypercholesterolemia in Brazil: hypercholesterolemia in Brazil: Identificationof seven novel LDLR gene Identificationof seven novel LDLR gene mutations.  mutations.  (2002)(2002)

Identification of Egr1 as the oncostatin M-Identification of Egr1 as the oncostatin M-induced transcription activatorthat binds to induced transcription activatorthat binds to sterol-independent regulatory element of sterol-independent regulatory element of human LDL receptor promoter.  human LDL receptor promoter.  (2002)(2002)

Sequencing of the coding exons of the Sequencing of the coding exons of the LRP1 and LDLR genes on individual DNA LRP1 and LDLR genes on individual DNA samples reveals novel mutations in both samples reveals novel mutations in both genes.  genes.  (2001)(2001)