Nagpal Neha, Agarwal Suneet
Division of Hematology/Oncology, Stem Cell Program, and Manton Center for Orphan Disease Research, Boston Children's Hospital, Boston, MA, USA.
Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
Nat Biomed Eng. 2025 Jun 27. doi: 10.1038/s41551-025-01429-1.
RNA engineering has yielded a new class of medicines but faces limitations depending on RNA size and function. Here we demonstrate the synthesis and enzymatic stabilization of telomerase RNA component (TERC), a therapeutically relevant long non-coding RNA (lncRNA) that extends telomere length and replicative lifespan in human stem cells. Compared with therapeutic mRNAs, engineered TERC RNA (eTERC) depends on avoiding nucleoside base modifications and incorporates a distinct trimethylguanosine 5' cap during in vitro transcription. We show that the non-canonical polymerase TENT4B can be repurposed to enzymatically stabilize synthetic RNAs of any size by catalysing self-limited 2'-O-methyladenosine tailing, which is critical for optimal eTERC function in cells. A single transient exposure to eTERC forestalls telomere-induced senescence in telomerase-deficient human cell lines and lengthens telomeres in induced pluripotent stem cells from nine patients carrying different mutations in telomere-maintenance genes, as well as primary CD34 blood stem/progenitor cells. Our results provide methods and proof of functional reconstitution for a stabilized, synthetic human lncRNA. eTERC may have therapeutic potential to safely extend replicative capacity in human stem cells.
RNA工程已经产生了一类新的药物，但根据RNA的大小和功能，仍面临一些局限性。在此，我们展示了端粒酶RNA组分（TERC）的合成及酶促稳定作用，TERC是一种与治疗相关的长链非编码RNA（lncRNA），可延长人类干细胞中的端粒长度和复制寿命。与治疗性mRNA相比，工程化TERC RNA（eTERC）需要避免核苷酸碱基修饰，并在体外转录过程中掺入一种独特的三甲基鸟苷5'帽结构。我们发现，非规范聚合酶TENT4B可通过催化自我限制的2'-O-甲基腺苷加尾，重新用于酶促稳定任何大小的合成RNA，这对eTERC在细胞中的最佳功能至关重要。单次短暂暴露于eTERC可防止端粒酶缺陷型人类细胞系中端粒诱导的衰老，并延长来自9名携带端粒维持基因不同突变的患者的诱导多能干细胞以及原代CD34血液干/祖细胞中的端粒。我们的结果为一种稳定的合成人类lncRNA提供了功能重建的方法和证据。eTERC可能具有安全延长人类干细胞复制能力的治疗潜力。
