be230cLecture9

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Unformatted text preview: right © 2001 Massachusetts Medical Society. Copyright © 2010 Massachusetts Medical Society. All rights reserved. HERE is concern that physicians are becom- The primary end point of the SOLVD treatment trial was death from any cause. The cause of death was also classified on standard forms after a review by the principal investigator at each center of the circumstances surrounding each death. Deaths from cardiovascular causes could be classified as due to pump failure, probable arrhythmia with some antecedent worsening of heart failure, or probable arrhythmia with no antecedent worsening of heart failure. As 10,11 2398 1G>C Mutation Result in a cryptic splice site and an immature transcript ! ! Matthew Stump, Quiming Gong Fig. RT-PCR analysis of mRNA isolated from IPS. A. Schematic illustrating normal splicing and the activation of a cryptic splice site within intron 9 by the G2398 +1G>C mutation (dashed arrow). Primers in exon 8 and exon 10 are indicated by arrows. Expected PCR products are shown below. B. mRNA was isolated from a normal IPS and the LQT2 mutant IPS. RT-PCR was performed using primers in exon 8 and exon 10. C. Sequence analysis of the 362 bp and 416 bp products from the LQT2 mutant IPS cells. The 416 bp product has a 54 bp insertion from intron 9. The arrow indicates the G2398+1G>C mutation. ! Patch clamp recording of LQT2 hiPSC ­CMs  ­40 mV  ­10 mV  ­90 mV  ­130 mV 50 µM TTX sensi1ve sodium current (INa) on day 27 Pacemaker current (If) on day 33 iPS Long QT Mutants have no detectable IKr Patch clamp recording of LQT2 hiPSC ­CMs 50 Figure 8: A, Functional expression of wild  ­40 type hERG channels in CHO cells. B, IKr recorded in early stage hESC-CMs (18-35 days post differentiation). Left, current blocked by 5 !M E4031 (indicated by arrows); right, voltage dependence of E4031-sensitive current. Stimulation protocol are shown above trace. ! Absence of IKr (no tail current CAVEATS: Often donʼt see IKr in Wild Type Cells Cecile Terronoire, George Wang, Jon Lu, Rocky Kass 26 hERG is Peri-nuclear in Long QT iPS Wild Type hES Long QT iPS Wild Type iPS Wild Type cel...
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