Intronic CRISPR Repair in a Preclinical Model of Noonan Syndrome–Associated Cardiomyopathy. Issue 11 (15th September 2020)

• Record Type: Journal Article
• Authors: Hanses, Ulrich; Kleinsorge, Mandy; Roos, Lennart; Yigit, Gökhan; Li, Yun; Barbarics, Boris; El-Battrawy, Ibrahim; Lan, Huan; Tiburcy, Malte; Hindmarsh, Robin; Lenz, Christof; Salinas, Gabriela; Diecke, Sebastian; Müller, Christian; Adham, Ibrahim; Altmüller, Janine; Nürnberg, Peter; Paul, Thomas; Zimmermann, Wolfram-Hubertus; Hasenfuss, Gerd; Wollnik, Bernd; Cyganek, Lukas
• Is Part Of: Circulation. Volume 142:Issue 11(2020)
• Publication Date: 2020-09-15
• Subjects: cardiomyopathy, hypertrophic -- clustered regularly interspaced short palindromic repeats -- gene editing -- induced pluripotent stem cells -- Noonan syndrome; Blood -- Circulation -- Periodicals; Cardiovascular system -- Periodicals; Cardiology -- Periodicals; Heart -- Diseases -- Periodicals; Blood Circulation; Cardiovascular System; Vascular Diseases; 616.1
• DOI: 10.1161/CIRCULATIONAHA.119.044794
• Languages: English
• ISSNs: 0009-7322

Abstract

Abstract : Background: Noonan syndrome (NS) is a multisystemic developmental disorder characterized by common, clinically variable symptoms, such as typical facial dysmorphisms, short stature, developmental delay, intellectual disability as well as cardiac hypertrophy. The underlying mechanism is a gain-of-function of the RAS–mitogen-activated protein kinase signaling pathway. However, our understanding of the pathophysiological alterations and mechanisms, especially of the associated cardiomyopathy, remains limited and effective therapeutic options are lacking. Methods: Here, we present a family with two siblings displaying an autosomal recessive form of NS with massive hypertrophic cardiomyopathy as clinically the most prevalent symptom caused by biallelic mutations within the leucine zipper-like transcription regulator 1 ( LZTR1 ). We generated induced pluripotent stem cell–derived cardiomyocytes of the affected siblings and investigated the patient-specific cardiomyocytes on the molecular and functional level. Results: Patients' induced pluripotent stem cell–derived cardiomyocytes recapitulated the hypertrophic phenotype and uncovered a so-far-not-described causal link between LZTR1 dysfunction, RAS–mitogen-activated protein kinase signaling hyperactivity, hypertrophic gene response and cellular hypertrophy. Calcium channel blockade and MEK inhibition could prevent some of the disease characteristics, providing a molecular underpinning for the clinical use of these drugs in patients with NS, but might not be a sustainable therapeutic option. In a proof-of-concept approach, we explored a clinically translatable intronic CRISPR (clustered regularly interspaced short palindromic repeats) repair and demonstrated a rescue of the hypertrophic phenotype. Conclusions: Our study revealed the human cardiac pathogenesis in patient-specific induced pluripotent stem cell–derived cardiomyocytes from NS patients carrying biallelic variants in LZTR1 and identified a unique disease-specific proteome signature. In addition, we identified the intronic CRISPR repair as a personalized and in our view clinically translatable therapeutic strategy to treat NS-associated hypertrophic cardiomyopathy. Abstract : Supplemental Digital Content is available in the text.