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<title>Targeted reduction of let-7a miRNA increases fetal hemoglobin in human adult erythroblasts | NIH Research Festival</title>
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<article><div class="banner banner--gray banner--compact" style="--banner-background-image: url('/sites/default/files/media/image/2021-03/2015.jpg');--banner-background-focus: 50% 50%"><div class="banner__wrapper"><div class="banner__container"><h2 class="banner__heading">NIH Research Festival</h2><div class="banner__content"><p><strong><time datetime="2015-09-16">September 16</time> – <time datetime="2015-09-18">18, 2015</time></strong></p></div></div></div></div></article>
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<div class="section"><div data-drupal-messages-fallback class="hidden"></div></div><main role="main"><div id="main-content" tabindex="-1"></div><div class="section"><div class="content-sidebar"><div class="content-sidebar__main"><div class="content-sidebar__main-content"><div id="block-mainpagecontent-2"><article><h1>Targeted reduction of let-7a miRNA increases fetal hemoglobin in human adult erythroblasts</h1><div class="schedule"><div class="schedule__label"><div class="schedule__label-text"><time datetime="2015-09-17">Thursday, September 17, 2015</time>
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– Poster Session III
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</div></div><div class="schedule__content"><div class="schedule__item"><div class="schedule__time">3:30 – 5:00 p.m.</div><div class="schedule__description"><p><strong>FAES Terrace</strong></p></div><div class="schedule__description"><p><strong>NIDDK</strong></p></div><div class="schedule__description"><p><strong>CELLBIO-6</strong></p></div></div></div></div><p><strong>FARE Award Winner</strong></p><h2>Authors</h2><ul><li>JF de Vasconcellos</li><li>C Byrnes</li><li>YT Lee</li><li>M Kaushal</li><li>JM Allwardt</li><li>A Rabel</li><li>JL Miller</li></ul><h2>Abstract</h2><p>Induction of fetal hemoglobin (HbF) has therapeutic importance for patients with hemoglobinopathies. Previous studies showed that let-7 miRNAs are regulated during the fetal-to-adult transition in the erythroid lineage and reduced expression of let-7 in adult CD34+ cells increased HbF levels; however these studies did not address the potential for targeting an individual let-7 family member to regulate HbF levels. We initially determined the expression levels of the let-7 miRNAs in purified cell populations sorted from human peripheral blood and let-7a was identified as a predominantly expressed species in reticulocytes. Therefore, we hypothesized that specifically targeting let-7a may be sufficient to regulate HbF. For this purpose, a lentiviral construct with the tough decoy (TuD) design to target let-7a was compared with empty vector controls. Transductions were performed in human CD34+ cells from five adult healthy volunteers cultivated ex vivo in erythropoietin-supplemented serum-free media for 21 days. Down-regulation of let-7a was confirmed by Q-RT-PCR at day 14 (control: 1.4E+07 +/- 2.4E+06 copies/ng; let-7a-TuD: 1.6E+06 +/- 4.6E+05 copies/ng; p=0.0003). Expression levels of globin genes were evaluated with significant increases in gamma-globin mRNA expression levels after let-7a-TuD transduction (control: 1.2E+06 +/- 6.8E+05 copies/ng; let-7a-TuD: 1.1E+07 +/- 4.5E+06 copies/ng; p=0.004). Remarkably, HPLC analyses at day 21 demonstrated robust increases in HbF levels after let-7a-TuD transduction (control: 4.7 +/- 0.6%; let-7a-TuD: 38.2 +/- 3.8%; p=0.00003). Our findings identify let-7a as a predominantly expressed let-7 family member in human reticulocytes and that targeted reduction of let-7a in erythroblasts is sufficient to cause robust increases in HbF levels.</p><p><strong>Scientific Focus Area</strong>: Cell Biology</p><p class="updated"><small>This page was last updated on Friday, March 26, 2021</small></p></article>
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