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I would really like to know 🙂 we do not have the exact mechanism which is crucial for better understanding. My guess is that the unfolded protein response (Ed Figure 6 a-b in the manuscript) is somehow connected. But we don't have a definitive answer yet.

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It's about spatial distribution. TENT5A is anchored to the ER membrane, thus being close to the RNAs being translated there. In our previous works we were unable to find any specific sequence motifs in TENT5 mRNA substrates.

I'm grateful for this milestone in my career - an experience that’s shaped me both scientifically and personally. Looking ahead with excitement to the next chapter and new opportunities to grow.

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🙏 Huge thanks to everyone involved - our incredible team (DziembowskiLab @iimcb.bsky.social), collaborators (University of #Warsaw #UW, Medical University of Warsaw #WUM, #IBB #PAS), and funding agencies (@erc.europa.eu, @ncn.gov.pl, #FNP, #PORT #Łukasiewicz). (20/n)

🇵🇱 Notably, this represents a landmark achievement as likely the first @nature.com article of the 21st century conducted entirely in #Polish laboratories! (19/n) #PolishScience

This publication represents the culmination of a long,challenging journey.We initially posted our findings as a #preprint on @biorxivpreprint.bsky.social in December 2022.The rigorous review process demanded additional experiments,which significantly strengthened our final manuscript.(18/n)

These findings have immediate implications for therapeutic mRNA design: optimization of ER-targeting signals could enhance TENT5A-mediated re-adenylation, potentially improving vaccine efficacy and reducing dosage requirements. (17/n)

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Notably, this effect was specific to #mRNA vaccines - #TENT5A deficiency did not impair antibody production in response to the protein-based #vaccine #Nuvaxovid (@novavax.bsky.social). (16/n)

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🐭 In vivo studies showed that TENT5A-deficient mice produced significantly less antigen-specific #antibodies following #mRNA #vaccination. This highlights the essential role of #TENT5A in mounting an effective immune response to mRNA-based vaccines. (15/n)

Our findings reveal a mechanistic principle: spatial proximity to ER-resident #TENT5A governs re-adenylation efficiency. This positions the #endoplasmic #reticulum as a central hub not only for #mRNA translation, but also for post-transcriptional regulation. (14/n)

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Surprisingly, comparative analysis revealed differential re-adenylation efficiency between vaccine formulations: @biontech.bsky.social #Pfizer #BNT162b2 demonstrated reduced re-adenylation compared to mRNA-1273, correlating with diminished #ER membrane association. (13/n)

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The re-adenylation phenomenon exhibits substrate specificity - consistently observed in synthetic mRNAs encoding proteins targeted to the #endoplasmic #reticulum (ER), including #spike #glycoprotein, #Zika virus antigens, and #malaria parasitic proteins. (12/n)

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We found that #macrophages—not dendritic cells—are the primary cells that take up mRNA vaccine #LNPs and express #TENT5A, positioning them as key players in regulating mRNA stability via re-adenylation. (11/n)

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We identified #TENT5A -a polyA polymerase our lab has studied extensively - as responsible for this effect. Its expression is induced by vaccine delivery, most likely through innate immune signaling (please compare our previous paper: doi.org/10.1126/scia...). #InnateImmunity(9/n)

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We then detected the same #polyA elongation pattern in #macrophages - both those sorted from muscles surrounding injection sites and in the in vitro models (mouse bone marrow-derived macrophages (mBMDMs) and human monocyte-derived macrophages (hMDMs)). (8/n)

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Crucially, when examining #mRNA #vaccines at injection sites in #mice, we discovered an unexpected phenomenon: #polyA tails were extended from ~100 to ~200 nucleotides - contradicting the expected degradation pattern observed in standard cell lines. (7/n) #mRNAStability

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In typical cellular models (HEK293T), we observed that mRNA-1273 degradation is initiated via removal of the terminal pentamer (mΨCmΨAG), followed by #CCR4-NOT-mediated #deadenylation—the canonical #mRNA #decay pathway. (6/n) #RNAMetabolism

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The #Moderna #mRNA-1273 vaccine possesses a ~100 nucleotide #polyA tail terminated with a distinctive pentamer (mΨCmΨAG), which served as a critical molecular marker, facilitating differentiation between processed and unprocessed tails. (5/n)

To overcome this obstacle, I developed a Dynamic Time Warping approach to detect and analyze vaccine molecules using #DRS with high fidelity, enabling precise characterization of #polyA tails from #therapeutic #mRNAs. (4/n)

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We employed @nanoporetech.com #direct #RNA #sequencing (DRS) to characterize individual therapeutic mRNA molecules - specifically focusing on #polyA #tail dynamics. However, #methylpseudouridine (mΨ) modifications in therapeutic mRNAs create significant analytical challenges. (3/n)

Our research elucidates a critical post-transcriptional modification enhancing #mRNA #vaccine efficacy. We've identified #TENT5A-mediated re-adenylation as a key determinant of therapeutic mRNA stability and immunogenicity in #SARS_CoV_2 #vaccines. (2/n) #mRNABiology

iimcb.bsky.social @iimcb.bsky.social · 16/04/2025

🎉 Incredibly proud to share my first-author paper in @nature.com! Heartfelt thanks to my mentor @andrzejdziembowski.bsky.social and the brilliant team at DziembowskiLab at @iimcb.bsky.social. More details in the 🧵below ⬇️

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Reposted by Paweł Krawczyk

🌐Our @nature.com paper is out! Researchers from IIMCB and other Polish institutions show how TENT5A boosts mRNA vaccine efficacy by stabilizing therapeutic mRNAs and enhancing antigen production. A step toward better therapies for cancer and beyond! 🧬 👉Read the story: bit.ly/3XYvQkI #Nature #mRNA

bit.ly

IIMCB

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Paweł Krawczyk @pakraw.bsky.social · 19/03/2025

The latest achievement from the Dziembowski Lab @IIMCB now published at @naturecomms.bsky.social. Ninetails is a #neural-network based tool using @nanoporetech.com #directRNA data to identify and quantify non-adenosines in #mRNA #polyA tails. Huge congrats to Natalia!

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Paweł Krawczyk @pakraw.bsky.social · 28/02/2025

My first (and probably the last), first-author @nature.com paper formally accepted. Huge effort of all people involved, mostly from the Laboratory of RNA Biology, IIMCB in Warsaw (all authors are from Polish institutions!). More details when the manuscript will be in press.

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Paweł Krawczyk @pakraw.bsky.social · 07/01/2025

Only 3 sentences... But we are almost there 😁

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Paweł Krawczyk @pakraw.bsky.social · 31/12/2024

Update - review received today. At least the reviewer managed to send it in the current year... hopefully there are nice wishes for the New Year