Структурно-функциональный анализ транскриптома Mycobacterium tuberculosis при развитии инфекции in vivo

Выводы.

1. Разработан новый универсальный метод анализа транскриптомов внутриклеточных патогенов из пораженной ткани организма хозяина, основанный на обогащении и последующем массированном секвенировании полученного набора бактериальной кДНК. Разработаны биоинформатические и статистические подходы к обработке полнотранскриптомных данных. Высокая чувствительность и специфичность метода продемонстрирована на примере анализа транскриптома М. tuberculosis в хронической фазе инфекции.

2. С помощью разработанного метода проведен анализ набора последовательностей M. tuberculosis H37Rv, транскрибирующихся в инфицированных тканях мышей, чувствительных и резистентных к инфекции, на 4-й и 6-й неделях развития болезни. В целом, структура транскриптома М. tuberculosis при развитии инфекции свидетельствует об активизации липидного метаболизма, метаболизма аминокислот, переходе к анаэробному дыханию, повышении экспрессии факторов модуляции иммунного ответа.

3. По результатам сравнительного анализа транскриптомов определено 209 генов, экспрессия которых повышается в ходе развития инфекции и не зависит от генетических особенностей организма-хозяина. Наибольший интерес представляют гены, относящиеся к функциональным категориям липидного метаболизма и клеточной стенки и клеточных процессов.

Заключение

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В данной работе впервые проведен параллельный широкомасштабный анализ транскриптомов М. tuberculosis в системе in vivo с использованием технологий секвенирования нового поколения. Получены данные о последовательностях М. tuberculosis, транскрибирующихся в легочных тканях генетически устойчивых и чувствительных к туберкулезу мышей в различных временных точках, что дало возможность установить факторы, ответственные за осуществление адаптации патогена к условиям конкретного микроокружения. Полученные транскриптомы М. tuberculosis были охарактеризованы качественно и количественно, выявлен ряд генов, уровень экспрессии которых инвариантно повышен при развитии инфекции. Экспрессия генов из этого ряда может рассматриваться как универсальная реакция микобактерий на различные стрессовые факторы внешней среды.

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