7.

012 Recitation 20 - 2011

Summarlz of Lecture 33: Familial Hypercholesterolemia (FH): If one of your arteries has a block in it, then blood flow is blocked and blood cannot get to the cells accessed by that artery. Plaques of cholesterol that build up in arteries restrict blood flow. Cholesterol contributes to the formation of plaques on blood vessel walls; this is called atherosclerosis. A heart attack occurs when there is blockage of blood vessels in the heart muscle, and a stroke occurs when there is blockage of blood vessels in brain. One gets cholesterol both from one's diet and from endogenous synthesis that involves a pathway in *hich the critical enzyme is called HMG coA reductase. Cholesterol is then iranspo.ted through your blood packaged in particles called LDL particles. Most cholesterol travels to your liver, which uses it to make bile acids. When the cholesterol arrives at your liver, the LDL particle is taken up by a cell-surface protein called the LDL receptor. People with mutations in the LDL teceptoi cannot take up cholesterol, and thus have very high levels of -These people have a disorder called FH. People with high cholesterol cholesterol in their blood. can be treated in four ways: they ian reduce their cholesterol intake, they,can take bile-acid binding resins, they can take drugs that inhibit HMG CoA reductase, or they could theoretically receive the wild-type LDL receptor gene through gene therapy'
You are investigating a method of gene therapy directed to the liver of a patient with familial hypercholesterolemil (FH). The pitient, who is homozygous for the FH alleles, is a human recipient for FH gene therapy. She suffered a massive heart attack with permanent damage at the age of 16, revealing the sev-rity of her disease. The procedure for her treatment is as follows. Fifteen percent of ihe patient's liver was removed. The liver tissue sample was then dissociated into individual hepatocytes (liver cells) and cultured in dishes. The cultured cells were provided with a wild-type cbpy of the gene that is mutated in this patient. The genetically altered hepatocytes fere injected baik into the patient via a vein entering the liver. After some time, the patient was given bile acid binding resins.
a) Why are the patient's liver cells specifically being targeted?

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7.0L2 Recitation 21 - 2011

Summary of Lectures 34 & 35:

Molecular evolution: Phylogeny is a branch of biology that explains how are organisms related to one another. The traditional way of determining how organisms are related to one another is to group them according to shared traits (phenotypes). However, another way is to look at the mutations in the coding and non-coding regions between the genome of different organisms: organisms that are closely related will have mutations predominantly in the non-coding regions and will have preserved coding regions since the mutations in the coding regions are deleterious. Over the course of time, due to random mutations, gene sequences randomly drift apart (diverge) unless sequence changes compromise fitness. Therefore, a comparable (homologous) DNA sequence in two organisms will be more divergent in more distantly related organisms. The noncoding sequences will diverge more rapidly than the coding sequences. One can also create an evolufionary tree based on comparative sequencing of 165 or 185 ribosomal RNA. The evolutionary distance between groups of organisms in the tree is proportional to the cumulative horizontal distance between the end of a branch and the node that joins the two groups.
The genetic distance of the human population across the globe can be measured by comparing the sequence of the mitochondria DNA that is always derived from the mother and has a minimum chance of recombination. Similarly the migration of males can be tracked by analyzing the Y chromosome.

Microarrays: Not all cancers respond equally well to all treatments, so knowing the specific type or subtype of cancer is important to successful treatment. Some types of cancers can be identified by histol,cgical tools, but other are best characterized by determining the gene expression profile. A DNA mlc.ou.tuy is a multiplex technology that allows comparison of cells based upon the expression of many different genes. A DNA microarray consists of an arrayed series of thousands of microscopic spots of DNA, each spot representing a gene. When a DNA microarray is probed with mRNA isolited from cells, individual mRNA molecules will hybridize to the appropriate DNA spot. If a gene is highly expressed, more mRNA will be made from that gene, so more mRNAwill hybridize to ihe coriesponding DNA spot, and the signal from that spot will be
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Recitation 20 Terms! Brian Hua R08/R09 Familial Hypercholesterolemia Cholesterol: A large hydrophobic organic molecule that plays an important role in cell membrane integrity but can also form buildups in the blood vessels causing dangerous impediments to blood circulation Plaque: A buildup of cholesterol crystals and dead cells in the blood vessel wall, which can impede blood circulation Atherosclerosis: A thickening of the blood vessel wall in response to plaques, further restricting blood circulation A heart attack can result if a blood vessel leading to the heart is blocked A stroke can result if a blood vessel leading to the brain is blocked HMG CoA reductase: A metabolic enzyme required in the synthesis of cholesterol in cells Low density lipoprotein (LDL): A packaged particle of cholesterol esters surrounded by a lipid monolayer that is used to transport cholesterol in the blood LDL receptor: A transmembrane protein receptor found notably on the liver cells that allows for the uptake of LDLs and hence cholesterol out of the blood stream, lowering blood cholesterol Familial hypercholesterolemia: A disease resulting the inability to take cholesterol out of the blood stream into the liver Typically a mutation in the LDL receptor gene that shows an incompletely dominant phenotype Gene therapy: A genetic therapeutic strategy to introduce ectopic genes into a diseased individual In familial hypercholesterolemia, gene therapy aims to introduce a WT copy of the LDL receptor gene so that functional LDL receptor can be expressed and LDL can be taken up by the liver and out of the blood Typically done by viral introduction of the WT gene Molecular Evolution Phylogeny: How biological organism are related to each other in the context of evolution Divergence: The events of the drifting two similar species to become more differentiated and unique

Evolutionary tree: A diagram tracing the phylogeny of organism, depicting the most related organism, as well as the points in time when divergence occurred and the ancestors of each organism 16S/18S ribosomal sequence: Genes that are common in all organisms across life that can be used to track evolution and divergence among organisms Mitochondrial DNA (mDNA): DNA that is passed on only by mothers that serves as a medium to trace the evolution and divergence of humans within a population Y-chromosome: DNA that is passed on from father to son that serves as a medium to trace the migration of males with a human population Microarrays Microarray: A biological technique used to assess the gene expression profile of particular cell types, tissues, or individuals Can be used to characterize different forms of cancers, as well as different levels of severity within the same cancer Directly measures mRNA levels, which is a direct readout of gene expression

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