Изогенная линия индуцированных плюрипотентных стволовых клеток ICGi036-A-1 от пациента с семейной гиперхолестеринемией, созданная путем коррекции патогенного варианта гена LDLR c.530C>T

Семейная гиперхолестеринемия является распространенным моногенным заболеванием, которое характеризуется повышенным содержанием холестерина в плазме крови, приводящим к хроническим заболеваниям сердечно-сосудистой системы с высоким риском и ранним проявлением развития патологий, вызванных атеросклеротическими поражениями кровеносных сосудов. Образование атеросклеротических бляшек при семейной гиперхолестеринемии в основном обусловлено патогенными вариантами гена рецептора липопротеинов низкой плотности LDLR (low-density lipoprotein receptor), играющего важную роль в метаболизме холестерина. В норме липопротеины низкой плотности, переносящие холестерин, связываются с рецептором LDLR на поверхности клеток печени и выводятся из кровотока путем интернализации гепатоцитами. При семейной гиперхолестеринемии происходит нарушение функционирования рецептора и значительное снижение интернализации липопротеинов низкой плотности. Это приводит к их накоплению в субэндотелиальном пространстве внутренней стенки кровеносных сосудов и вызывает атерогенез – образование атеросклеротических бляшек. На сегодняшний день не существует эффективных и универсальных подходов к диагностике и лечению семейной гиперхолестеринемии. Актуальным подходом для исследования молекулярно-генетических особенностей заболевания и разработки систем скрининга химических соединений – потенциальных лекарственных препаратов – является создание клеточных моделей на основе индуцированных плюрипотентных стволовых клеток (ИПСК) пациентов. Целью нашей работы было создание изогенной генетически модифицированной линии индуцированных плюрипотентных стволовых клеток путем коррекции патогенного аллельного варианта c.530C гена LDLR в линии ИПСК, полученной ранее от пациента-компаундной гетерозиготы с семейной гиперхолестеринемией. Созданная изогенная клеточная линия ИПСК отличается от исходной только одной скорректированной нуклеотидной заменой, что позволяет исследовать непосредственное влияние данного патогенного генетического варианта на физиологические изменения в релевантных дифференцированных клетках. Для коррекции однонуклеотидных замен использован CRISPR/Cas9-опосредованный метод редактирования оснований. Полученная генетически модифицированная линия ИПСК обладает свойствами плюрипотентности, имеет нормальный кариотип, идентичный исходной линии набор коротких тандемных повторов и может быть использована для формирования дифференцированных производных, необходимых при разработке релевантных клеточных моделей.

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