Laboratory of Molecular Mechanisms of Innate Immunity and Nucleic Acid Sensing
Referent: Prof. Anna Kajaste-Rudnitski
Co-workers: Cristina Belgiovine (Post-doc), Alessandro Mapelli (Research fellow), Radu Marian Rivara (Research fellow)
The AKR Lab studies the molecular mechanisms of host-vector interplay and innate immunity in the context of gene therapy and investigates these pathways in the context of pathological conditions of the central nervous systems and autoimmune diseases. The long-term goal is to provide insight into how pathogen recognition and consequent innate immune signalling may affect efficacy and safety of gene therapy approaches in clinically relevant target cells as well as to shed light on how these same pathways may contribute to autoimmune and inflammatory pathologies.
Research Topics:
Innate immunity and nucleic acid sensing are involved in an increasing number of biological processes from antiviral defences to tissue homeostasis and disease. Host antiviral factors and nucleic acid sensors play a pivotal role also in the efficacy and safety of genetic manipulation. On the other hand, the same sensors that protect us from viral infection and potentially hamper efficient gene engineering can also drive specific human autoimmune diseases when they are inappropriately activated by endogenous nucleic acids. On these premises, we study the molecular mechanisms of
host-vector interplay and innate immunity in the context of gene therapies in relevant target tissues such as the central nervous system and hematopoietic stem cells and investigate these pathways in the context of pathological conditions of the central nervous systems, autoimmune and inflammatory diseases. Together, our efforts will provide insight into how nucleic acid sensing and innate immune signalling may affect efficacy and safety of gene therapy approaches in clinically relevant target cells as well as to shed light on how these same pathways contribute to autoimmune and inflammatory pathologies.
- Innate Sensing of Gene Therapy Vectors
This area of research is focused on dissecting antiviral factors and innate sensing pathways in the context of genetic engineering. Building on our past work, we are addressing vector signalling and innate immune restriction across delivery platforms and in clinically relevant target cells. These basic studies of vector-host interactions will allow modulation of
identified host factors or innate sensing pathways in the context of transduction or gene editing with the goal of rendering gene engineering as inert as possible, while maximizing its efficiency.
- Nucleic Acid Immunity in Inflammatory Diseases
Here we aim to investigate innate immunity and nucleic acid sensing in the context of inflammatory diseases such as the Aicardi-Goutières Syndrome (AGS), a rare monogenic encephalopathy in which aberrant activation of innate sensing is thought to drive disease but the precise molecular mechanisms and cell types involved remain elusive. Among other systems, we use human induced pluripotent stem cells (iPSC) harbouring AGS loss of function alleles to dissect triggers of disease in cells of the central nervous system. These studies will provide insight into the pathological cascades causing disease informing the development of targeted therapies and of stealth gene engineering strategies that remain susceptible to similar mechanisms of innate sensing.
- Molecular Mechanisms of Virus-Host Interactions
This area of research is focused on dissecting the molecular mechanisms of antiviral restriction during viral entry and the role of these factors in hampering gene delivery relying on the same cellular pathways to access the cytosol. We are also addressing regulation and additional biological roles of these antiviral factors in the context of healthy and malignant tissues.
Main External Collaborators:
Angela Bachi (IFOM, MI, Italy)
Luigi Naldini (SR-Tiget, MI, Italy)
Alessandro Aiuti (SR-Tiget, MI, Italy)
Angelo Lombardo (SR-Tiget, MI, Italy)
Angela Gritti (SR-Tiget, MI, Italy)
Vasco Meneghini (SR-Tiget, MI, Italy)
Juan Bueren (CIEMAT, Madrid, Spain)
Federico Mingozzi (Spark Therapeutics, Philadelphia, USA)
Giuseppe Ronzitti (Genethon, Paris, France)
Angelo d’Alessandro (University of Colorado, Denver, USA)
Selected publications:
1. Costa-Verdera H, Meneghini V, Fitzpatrick Z, Abou Alezz M, Fabyanic E, Huang X, Dzhashiashvili Y, Ahiya A, Mangiameli E, Valeri E, Crivicich G, Piccolo S, Cuccovillo I, Caccia R, Chan YK, Bertin B, Ronzitti G, Engel EA, Merelli I, Mingozzi F, Gritti A, Kuranda K, Kajaste-Rudnitski A. AAV vectors trigger DNA damage response-dependent pro-inflammatory signalling in human iPSC-derived CNS models and mouse brain. Nat Commun. 2025 Apr 18;16(1):3694.
2. Valeri E, Breggion S, Barzaghi F, Abou Alezz M, Crivicich G, Pagani I, Forneris F, Sartirana C, Costantini M, Costi S, Marino A, Chiarotto E, Colavito D, Cimaz R, Merelli I, Vicenzi E, Aiuti A, Kajaste-Rudnitski A. A novel STING variant triggers endothelial toxicity and SAVI disease. J Exp Med. 2024 Sep 2;221(9):e20232167.
3. Valeri E, Unali G, Piras F, Abou-Alezz M, Pais G, Benedicenti F, Lidonnici MR, Cuccovillo I, Castiglioni I, Arévalo S, Spinozzi G, Merelli I, Behrendt R, Oo A, Kim B, Landau NR, Ferrari G, Montini E, Kajaste-Rudnitski A. Removal of innate immune barriers allows efficient transduction of quiescent human hematopoietic stem cells. Mol Ther. 2024 Jan 3;32(1):124-139.
4. Unali G, Crivicich G, Pagani I, Abou-Alezz M, Folchini F, Valeri E, Matafora V, Reisz JA, Giordano AMS, Cuccovillo I, Butta GM, Donnici L, D'Alessandro A, De Francesco R, Manganaro L, Cittaro D, Merelli I, Petrillo C, Bachi A, Vicenzi E, Kajaste-Rudnitski A. Interferon-inducible phospholipids govern IFITM3-dependent endosomal antiviral immunity. EMBO J. 2023 Mar 27:e112234.
5. Ferrari S, Jacob A, Cesana D, Laugel M, Beretta S, Varesi A, Unali G, Conti A, Canarutto D, Albano L, Calabria A, Vavassori V, Cipriani C, Castiello MC, Esposito S, Brombin C, Cugnata F, Adjla O, Ayuso E, Merelli I, Villa A, Di Micco R, Kajaste-Rudnitski A, Montini E, Penaud-Budloo M, Naldini L. Choice of template delivery mitigates the genotoxic risk and adverse impact of editing in human hematopoietic stem cells. Cell Stem Cell 2022 Oct 6;29(10):1428-1444.e9.
6. Giordano AMS, Luciani M, Gatto F, Abou Alezz M, Beghè C, della Volpe L, Migliara A, Valsoni S, Genua M, Dzieciatkowska M, Frati G, Tahraoui-Bories J, Giliani S, Orcesi S, Fazzi E, Ostuni R, D’Alessandro A, Di Micco R, Merelli I, Lombardo A, Reijns MAM, Gromak N, Gritti A, Kajaste-Rudnitski A. DNA damage contributes to neurotoxic inflammation in Aicardi-Goutières Syndrome astrocytes. J Exp Med. 2022 Apr 4;219(4):e20211121.
7. Soldi M, Sergi Sergi L, Unali G, Kerzel T, Cuccovillo I, Capasso P, Annoni A, Biffi M, Rancoita PMV, Cantore A, Lombardo A, Naldini L, Squadrito ML, Kajaste-Rudnitski A. Laboratory-Scale Lentiviral Vector Production and Purification for Enhanced Ex Vivo and In Vivo Genetic Engineering. Mol Ther Methods Clin Dev. 2020 Oct 20;19:411-425.
8. Piras F and Kajaste-Rudnitski A. Antiviral immunity and nucleic acid sensing in haematopoietic stem cell gene engineering. Gene Ther. 2020 Jul 13:1-13.
9. Petrillo C, Calabria A, Piras F, Capotondo A, Spinozzi G, Cuccovillo I, Benedicenti F, Naldini L, Montini E, Biffi A, Gentner B, Kajaste-Rudnitski A. Assessing the impact of Cyclosporine A on lentiviral transduction and preservation of human hematopoietic stem cells in clinically relevant ex-vivo gene therapy settings. Hum Gene Ther. 2019 Apr 30.
10. Petrillo C, Thorne LG, Unali G, Schiroli G, Giordano AMS, Piras F, Cuccovillo I, Petit SJ, Ahsan F, Noursadeghi M, Clare BS, Genovese P, Gentner B, Cittaro D, Naldini L, Towers GJ, Kajaste-Rudnitski A. Cyclosporine H Overcomes Innate Immune Restrictions to Improve Lentiviral Transduction and Gene Editing In Human Hematopoietic Stem Cells. Cell Stem Cell. 2018 Dec 6;23(6):820-832.e9. Recommended in F1000Prime as being of special significance in its field.
11. Piras F, Riba M, Petrillo C, Lazarevic D, Cuccovillo I, Bartolaccini S, Stupka E, Gentner B, Cittaro D, Naldini L, Kajaste-Rudnitski A. Lentiviral Vectors Escape Innate Sensing but Trigger p53 In Human Hematopoietic Stem and Progenitor Cells. EMBO Mol Med. Sep;9(9):1198-1211. 2017.
12. Petrillo C, Cesana D, Piras F, Bartolaccini S, Naldini L, Montini E, Kajaste-Rudnitski A. Cyclosporin A and Rapamycin relieve distinct lentiviral restriction blocks in hematopoietic stem and progenitor cells. Mol Ther., Feb. 23. 2015.