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:
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.
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 SC, 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. doi: 10.1084/jem.20211121.
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.
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.
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 Cyclosporin A on Lentiviral Transduction and Preservation of Human Hematopoietic Stem Cells in Clinically Relevant Ex Vivo Gene Therapy Settings. Hum Gene Ther. 2019 Sep;30(9):1133-1146. doi: 10.1089/hum.2019.016. Epub 2019 May 24.
Petrillo C., Thorne L., Unali G., Schiroli G., Giordano A. M. S., Piras F., Cuccovillo I., Petit S., Ahsan F. Noursadeghi M., Clare S., Genovese P., Gentner B., Naldini L., Towers G., Kajaste-Rudnitski A. Cyclosporine H overcomes IFITM3-mediated innate immune restriction to lentiviral transduction and gene editing in human hematopoietic stem cells. Cell Stem Cell, 2018 Dec 6;23(6):820-832.e9. doi:10.1016/j.stem.2018.10.008. Epub 2018 Nov 8.
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. 2017 Sep;9(9):1198-1211.
Zonari E, Desantis G, Petrillo C, Boccalatte FE, Lidonnici MR, Kajaste-Rudnitski A, Aiuti A, Ferrari G, Naldini L, Gentner B. Efficient ex vivo engineering and expansion of highly purified human hematopoietic stem and progenitor cell populations for gene therapy. Stem Cell Reports 2017 Apr 11;8(4):977-990.
Kajaste-Rudnitski A, Naldini L. Cellular innate immunity and restriction of viral infection: implications for lentiviral gene therapy in human hematopoietic cells. Hum Gene Ther. 2015 Apr;26(4):201-9.
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. 2015 Feb;23(2):352-62.
Escobar G, Moi D, Ranghetti A, Ozkal-Baydin P, Squadrito ML, Kajaste-Rudnitski A, Bondanza A, Gentner B, De Palma M, Mazzieri R, Naldini L. Genetic engineering of hematopoiesis for targeted IFN-α delivery inhibits breast cancer progression. Sci Transl Med. 2014. 6(217):217.
Di Pietro A*, Kajaste-Rudnitski A*, Oteiza A, Nicora L, Towers GJ, Mechti N, Vicenzi E*. TRIM22 inhibits influenza A virus infection by targeting the viral nucleoprotein for degradation. J Virol. 2013 Apr; 87(8):4523-4533. (* shared first authorship)
Kajaste-Rudnitski A*, Marelli S, Pultrone C, Pertel T, Mechti N, Poli G, Luban J, Vicenzi E. TRIM22 Inhibits HIV-1 Transcription independently of its E3-ubiquitin ligase activity, Tat and NF-{kappa}B responsive LTR elements. J Virol. 2011. 85(10):5183-96.
(* corresponding authorship)
Kajaste-Rudnitski A, Galli L, Nozza S, Tambussi G, Di Pietro A, Pellicciotta G, Monti A, Mascagni P, Moro M, Vicenzi E. Induction of protective antibody response by MF59- adjuvanted 2009 pandemic A/ H1N1v influenza vaccine in HIV-1 infected individuals. AIDS 2011. 25(2):177-83.
Kajaste-Rudnitski A, Mashimo T, Frenkiel MP, Guenet JL, Lucas M, Despres P. The 2′, 5′ oligoadenylate synthetase 1b is a potent inhibitor of West Nile virus replication inside infected cells. J Biol Chem. 2006. 281:4624-37.