Laura CROFT, PhD
Senior Research Fellow, Institute of Health and Biomedical Innovation, School of Biomedical Science, Queensland University of Technology, Australia
Team Leader Drug Development, Cancer and Ageing Research Program, Queensland University of Technology, Australia
Dr Croft received her Bachelor of Science degree from Griffith University, Brisbane, Australia in 2001, and continued her doctoral studies from 2002 to 2006 at Griffith University and at the Queensland Institute of Medical Research, Brisbane, Australia. Dr Croft’s doctoral studies focused on selenoprotein translation mechanisms, and selenium biology and metabolism. Dr Croft also spent one year of her PhD at the Karolinska Institute, Stockholm, Sweden to study redox regulation of protein function in Prof Arne Holmgren’s lab. During her post-doctoral position at QIMR, 2006-2011, Dr Croft started utilising antisense oligonucleotides as a means to alter splicing and study protein function. In 2012, Dr Croft joined the Cancer and Ageing Research Program (CARP) at Queensland University of Technology (QUT) where her research focus became targeting DNA repair proteins in cancer using oligonucleotides and translating basic research into novel cancer therapeutics. Dr Croft is currently developing novel, nucleic acid-based cancer therapies by targeting DNA repair pathways. Dr Croft is team leader of drug development within CARP at QUT.
Targeting DNA repair pathways in cancer using oligonucleotides
Genome stability pathways offer a promising area of therapeutic intervention in cancer. Specific inhibitors of key DNA repair proteins have shown promising results as combination therapies and more recently also as single agents. We use nucleic acid-based technology to target DNA repair proteins in cancer and explore this technology as a potential cancer therapeutic. We show that specific oligonucleotides inhibit DNA repair, induce DNA damage, genomic fragmentation and apoptosis in multiple cancer cell lines, and suppress tumour growth in mouse xenograft models of prostate and lung cancer without any apparent toxicity to the animals.
Housheng Hansen HE, PhD
Associate Professor, Department of Medical
Biophysics, University of Toronto, Canada
Senior Scientist, Princess Margaret Cancer Centre,
University Health Network, Canada
Dr. He received his B.S. degree in physics from Beijing Normal University in the year of 2003, and then transitioned to the genomics and noncoding RNA fields for his Ph.D training in the Institute of Biophysics, Chinese Academy of Sciences. He received his postdoctoral training in cancer genomics and epigenomics at Dana-Farber Cancer Institute. After completing his postdoctoral training in 2011, Dr. He worked as an instructor at Harvard Medical School and was recruited as a scientist at the Princess Margaret Cancer Centre in 2013.
The He laboratory applies a variety of genomic, epigenomic experimental and computational approaches to elucidate the functional role of epigenetic regulation in cancer development, progression and drug response/resistance, with a special focus on the interplay between epigenetic regulator and noncoding RNA under stress conditions such as hypoxia. Dr. He’s work in the fields of noncoding RNA and epigenetics has resulted in ~50 peer-reviewed publications in high-impact journals including Nature, Cell, Nature Genetics, Nature Methods, Cancer Cell and Genome Research.
Professor, Institute for Genetic Medicine
Hokkaido University, Japan
Dr. Hirose was awarded his Ph.D. at Nagoya University in 1995 under supervision of Prof. Masahiro Sugiura, and stayed in Sugiura lab to study the mechanism of chloroplast RNA editing as research assistant professor of Nagoya University. In 1999, Dr. Hirose moved to Prof. Joan Steitz laboratory in Yale University School of Medicine as the HFSP fellow where he worked on the mechanism of intronic snoRNA biogenesis in mammalian cells. In 2004, he moved back to Japan to start his own research group as associate professor at Tokyo Medical & Dental University (until 2005) and then as group leader at National Institute for Advanced Industrial Science and Technology. In 2013, he joined Institute of Genetic Medicine (IGM), Hokkaido University as Full Professor. Since 2016, he has been appointed as the Vice-director of IGM. Dr. Hirose discovered a new class of long noncoding RNA (lncRNA) called “architectural RNAs” that act as structural scaffolds of subcellular granules. The ultimate goal of his research is to establish the novel genetic rule underlying the function of lncRNAs and apply it to develop new therapeutic tools.
Nuclear architecture by long noncoding RNAs through phase separation
Specific long noncoding RNAs have been identified as the structural scaffold of distinct membraneless nuclear bodies, therefore termed as architectural RNAs (arcRNAs). The arcRNAs build massive nuclear bodies by sequestrating multiple RNA-binding proteins possessing intrinsically disordered regions (IDRs). This process likely proceeds via liquid-liquid phase separation (LLPS) where arcRNA locally concentrates IDR containing RNA binding proteins. I will discuss the molecular mechanism of arcRNAs function revealed by identifying functional elements embedded in the arcRNA sequences as well as interacting RNA-binding proteins that induce LLPS.
Chandrasekhar KANDURI, PhD
Professor in RNA Biology
Department of Medical Biochemistry and Cell Biology
University of Gothenburg, Sweden
Dr. Chandra Kanduri received his PhD from Banaras Hindu University, India, in 1997. For postdoctoral training, he moved to Prof Rolf Ohlsson’s lab at Uppsala University, Sweden, where he explored the functional role of chromatin insulator protein CTCF in genomic imprinting using H19 and IGF2 imprinted cluster as a model system. He later continued his work on long noncoding RNA Kcnq1ot1 and genomic imprinting as an independent investigator. His work, as an early career scientist, contributed significantly towards understanding the functional interaction between RNA and chromatin. His further work on RNA-chromatin connection, revealed RNA molecules enriched in the active and inactive chromatin regions, and also possible mechanisms by which chromatin enriched lncRNAs targeted across the genome.
He became full professor of RNA biology at Uppsala university in 2010. In 2012, he moved his research activities to University of Gothenburg. His current research primarily focused on developing novel long noncoding RNAs (lncRNA) based strategies for the treatment of drug resistant and difficult to treat cancers through exploiting cell cycle based functional screens.
Functional and mechanistic roles of long noncoding RNAs (lncRNAs) in cancer development and progression
Despite rapid improvements in unveiling the importance of long noncoding RNAs (lncRNA) in all aspects of cancer biology, there is still a lot of void in how lncRNAs mechanistically influence various biological processes associated with cancer development. Towards this, by using RNA-seq data from cell cycle-based high-throughput functional screens and different neuroblastoma (NB) prognostic sub-groups, we have identified several cancer-relevant, therapeutically applicable lncRNAs. Functional investigations of these lncRNAs revealed their nexus with oncogenic and tumor-suppressor pathways. I will discuss how these cancer relevant lncRNAs can be used as potential therapeutic targets in different cancer types.
Ailong KE, PhD
Professor, Department of Molecular Biology and Genetics,
Cornell University, US
Dr. Ailong Ke received his B.S. from the University of Science and Technology of China in 1995, and Ph.D. in Biophysics with Cynthia Wolberger from the Johns Hopkins University School of Medicine in 2002. After his post-doctoral training with Jennifer Doudna at UC Berkeley, he started as an Assistant Professor at Cornell University in 2005, and rose to the full Professorship in 2017. Since independence, he has been working in the areas of RNA 3’-end processing and degradation, metabolite-sensing riboswitches, and more recently, the CRISPR-Cas immunity system. He has published over 40 papers in journals such as Nature, Science, Cell, Molecular cell, NSMB, PNAS, and RNA.
Spacer acquisition mechanism in type II-A CRISPR system.
Molecular memory is created when a short foreign DNA-derived prespacer is integrated into the CRISPR array as a new spacer. Whereas the RNA-guided CRISPR interference mechanism varies widely among CRISPR-Cas systems, the spacer integration mechanism is essentially identical. The conserved Cas1 and Cas2 proteins form an integrase complex consisting of two distal Cas1 dimers bridged by a Cas2 dimer. The prespacer is bound by Cas1-Cas2 as a dual-forked DNA, and the terminal 3′-OH of each 3′ overhang serves as an attacking nucleophile during integration. The prespacer is preferentially integrated into the leader-proximal region of the CRISPR array, guided by the leader sequence and a pair of inverted repeats inside the CRISPR repeat. Spacer integration in the well-studied Escherichia coli type I-E CRISPR system also relies on the bacterial integration host factor. In type II-A CRISPR, however, Cas1-Cas2 alone integrates spacers efficiently in vitro; other Cas proteins (such as Cas9 and Csn2) have accessory roles in the biogenesis phase of prespacers. I will present our structural and biochemical efforts in revealing the spacer acquisition mechanism in the Enterococcus faecalis type II-A CRISPR system.
Ekkehard LEBERER, PhD
Professor, Senior Director, Sanofi, Germany
Founding Director, COMPACT Consortium
Since joining Hoechst Marion Roussel in 1998, Dr. Leberer carried out various managing roles in this company, Sanofi’s predecessor companies and Sanofi itself, including responsibilities in functional genomics, biological sciences and external innovation for oligonucleotide-based therapeutics. He has also served as Head of Biotechnology Germany and a member of the Scientific Review Committee of Aventis Pharma Germany.
Prior to joining pharmaceutical industry, Dr. Leberer served as Senior Research Officer in genetics and genomics at the Biotechnology Research Institute, National Research Council of Canada, Montreal. His research has focused on the molecular mechanisms of signal transduction and the role of signalling molecules in human diseases. He is the principal discoverer of the p21 activated protein kinase (PAK) family of cell signalling proteins and of novel virulence-inducing genes in pathogenic fungi. He is co-author of more than 60 publications in prestigious peer-reviewed journals including Nature and Science.
Delivering Therapeutic Oligonucleotides across Biobarriers: Opportunities and Challenges in Drug Development
• Macromolecular biopharmaceuticals such as oligonucleotides have a huge pharmacological potential but their widespread therapeutic application has been very limited due to pharmacokinetic and drug disposition limitations at both the tissue and cellular level.
• The presentation will address these delivery limitations and summarize the work of a European consortium (www.compact-research.org) of pharma companies and academic partners to improve nanocarrier-based delivery technologies that can overcome these limitations.
Dong-ki LEE, PhD
Professor of Chemistry, Sungkyunkwan University, Korea
Founder and CEO, OliX Pharmaceuticals, Korea
Prof. Dong-ki Lee received B. S. in Chemistry from Korea Advanced Institute of Science and Technology (KAIST) in 1993, and Ph. D. in Biochemistry from Cornell University in 1999. After post-doctoral training in Pohang University of Science and Technology (POSTECH), Toolgen Inc., and KAIST, he joined POSTECH as an assistant professor in 2004. In 2008, He moved to Sungkyunkwan University and is a full professor of Chemistry since 2012. In 2008, Prof. Lee was selected as the principal investigator of the Global Research Laboratory program, funded by Korean government, to develop novel RNAi medicine. He is currently serving as the Asian editor of “Nucleic Acid Therapeutics” and a editorial board member of “Molecular Therapy: Nucleic Acids”. His work on novel siRNA structures, nucleic acid aptamers, and eukaryotic gene regulation has been published as over 80 papers including prestigious journals such as Nature, Cell, PNAS, and Molecular Therapy. In 2010, he founded OliX Pharmaceuticals, a RNAi therapeutics company focusing on topically administrable diseases, and serves as Chief Executive Officer. OliX successfully went public in July 2018, with 370M USD valuation.
Asymmetric siRNA Targeting Fibrotic and Ocular Disorders
OLX10010, an anti-fibrotic cell-penetrating asymmetric siRNA (cp-asiRNA) targeting connective tissue growth factor (CTGF), effectively reduces target gene expression as well as expression of fibrotic markers in animal model study. Preclinical as well as clinical study update of OLX10010 in anti-skin scar will be presented. In addition to skin scar, OLX10010 has a potential to be developed as therapeutics targeting various fibrotic disorders. We will present preclinical study data of OLX10010 in other fibrotic diseases in lung and eye, such as idiopathic pulmonary fibrosis (IPF) and subretinal fibrosis.
Wei LI, PhD
Professor of Bioinformatics,
Dan L. Duncan Cancer Center
Department of Molecular and Cellular Biology
Baylor College of Medicine, USA
Dr. Wei Li is a Professor of Bioinformatics in the Dan L. Duncan Cancer Center at Baylor College of Medicine. He received his PhD in Bioinformatics from the Chinese Academy of Sciences (2003), and was an Associate Director of Bioinformatics at Beijing Genomics Institute (BGI; 2002-2004). After his postdoctoral training with Dr. Xiaole Shirley Liu in the Department of Biostatistics and Computational Biology at Harvard (2004-2007), he was recruited to Baylor as an Assistant Professor in 2007 and was promoted to tenured Full Professor in 2016. His research is focused on the design and application of bioinformatics algorithms to elucidate global epigenetic mechanisms in normal development and diseases, such as cancer. He has a solid track record in developing widely used open-source bioinformatics software, such as MACS (~5,000 citations) for ChIP-seq. Since establishing his own bioinformatics lab, he has (as of April 2018) (1) Published ~120 peer-reviewed papers through solid methodology development and extensive collaborative research, including 19 senior-author papers in Nature and Cell series. (2) Been well-funded with total external funding >$1.0 million per year, including 4 PI grants from NIH and Texas CPRIT. (3) Mentored the first 6 postdoc trainees to start their tenure-track faculty positions in the US. Dr. Li received many prestigious awards, including the New Investigator Award from Department of Defense (2010), and the Michael E. DeBakey Excellence in Research Award (2016).
Alternative polyadenylation (APA) is emerging as a pervasive mechanism in the regulation of more than 70% of human genes. The role of APA in human cancer is largely overlooked due to the lack of genome-wide APA profiling on large clinical cohorts. To overcome this limitation, we recently developed DaPars algorithm (Nature Communications 2014) for the de novo identification of APA from standard RNA-seq. Through DaPars analyses of thousands of TCGA tumor RNA-seq data, we revealed ~1,300 highly recurrent 3’UTR shortening genes, nominated CFIm25 (Nature 2014) as a master regulator of 3`-UTR shortening that links APA to glioblastoma tumor suppression, and suggested a surprising enrichment of 3’UTR shortening among transcripts that are predicted to act as competing-endogenous RNAs (ceRNAs) for tumor suppressor genes (Nature Genetics 2018).
Zhen LI, PhD
Senior Vice President, Chemistry and Non-Clinical Development
Arrowhead Pharmaceuticals, USA
Dr. Zhen Li is Senior Vice President, Chemistry and Non-Clinical Development at Arrowhead Pharmaceuticals. She leads discovery chemistry, biology and non-clinical development at Arrowhead. She has led the discovery and development of Arrowhead TRiMTM platform for RNAi based therapeutics for hepatic and non-hepatic targets, and she is the key inventor of the platform. Prior to joining Arrowhead in 2014, she held leadership positions at Merck, Schering-Plough and Novartis, and led teams in drug discovery and process development in small molecule pharmaceuticals as well as RNAi therapeutics covering a range of disease areas. She has a demonstrated track record in leading multiple drug development programs from early to late stage. Dr. Zhen Li received her Bachelor of Science degree from Peking University and her Ph.D. from Harvard University.
TRiMTM Platform Based RNAi Therapeutics in Treating HBV and Cardiometabolic Diseases
One of the more important recent advances in biology, RNA interference is a natural cellular process where short oligonucleotides can be used to silence gene expression and regulate the production of proteins. Arrowhead’s RNAi-based therapeutics leverage this natural pathway to shut down specific genes that cause disease.
The presentation will focus on Arrowhead’s clinical candidate ARO-HBV for the treatment of Hepatitis B Virus infection. HBV infection presents a huge unmet medical need, as it affects over 250 million people worldwide and currently there is no cure for this devastating disease. In the presentation, some of our earliest clinical data on ARO-HBV will be presented, and the journey in discovering this powerful drug candidate will be discussed. The presentation will also discuss Arrowhead’s preclinical candidates ARO-ANG3 and ARO-APOC3 for the treatment of cardiometabolic disease such as hypertriglyceridemia. Gene knockdown, reduction of protein levels, and reduction of triglycerides in disease models for Arrowhead’s preclinical candidates ARO-ANG3 and ARO-APOC3 will be presented.
Muthiah MANOHARAN, PhD
Senior Vice President, Drug Discovery, Alnylam Pharmaceuticals, USA
Board Director, Oligonucleotide Therapeutics Society
Dr. Muthiah (Mano) Manoharan serves as a Senior Vice President and a Distinguished Research Scientist at Alnylam Pharmaceuticals, Cambridge, Massachusetts, USA. Dr. Manoharan joined Alnylam in 2003. He built the chemistry group at Alnylam and pioneered the discovery and development of RNA interference-based human therapeutics. Dr. Manoharan has had a distinguished career as a world-leading chemist in the areas of oligonucleotide chemical modifications, conjugation chemistry, and delivery platforms (lipid nanoparticles, polymer conjugates, and complex-forming strategies). He is an author of more than 200 publications (nearly 37,000 citations with an h-index of 85 and an i10-index of 329) and over 400 abstracts, as well as the inventor of over 225 issued U.S. patents.
Advances in RNAi Therapeutics Using Chemistry
RNA interference (RNAi) based therapeutics has evolved for the past two decades, and this year finally there is one approved siRNA-based drug representing the success of RNAi mechanisms of action: ONPATTRO or patisiran. On celebrating this occasion, we will review the key milestones of the past 16 years of drug discovery at Alnylam. This includes
• Chemical modifications, their functions, and pharmacological properties
• Delivery platforms (lipid nanoparticles and conjugates)
• Mechanisms of Delivery of RNAi therapeutics targeted to liver
• Emerging new approaches of overcoming RNAi-mediated, hybridization based off-target effects
• Use of Reversirs to modulate the RNAi effects
Craig MELLO, PhD
Professor, University of Massachusetts
Howard Hughes Medical Institute, USA
Nobel Prize in Physiology or Medicine (2006)
Dr. Mello received his BS degree in Biochemistry from Brown University in 1982, and PhD from Harvard University in 1990. From 1990 to 1994, he conducted postdoctoral research at the Fred Hutchinson Cancer Research Center in Seattle, WA. Now Dr. Mello is an Investigator of the Howard Hughes Medical Institute, the Blais University Chair in Molecular Medicine and Co-director of the RNA Therapeutics Institute at the University of Massachusetts Medical School.
Besides the Nobel Prize, Dr. Mello’s work was recognized with numerous awards and honors, including the National Academy of Sciences Molecular Biology Award (2003), the Wiley Prize in Biomedical Sciences from Rockefeller University (2003), Brandeis University’s Lewis S. Rosnstiel Award for Distinguished Work in Medical Research (2005), the Gairdner Foundation International Award (2005), the Massry Prize (2005), the Paul Ehrlich and Ludwig Darmstaedter Award (2006), the Dr. Paul Janssen Award for Biomedical Research (2006), the Hope Funds Award of Excellence in Basic Research (2008). He is a member of the National Academy of Sciences, the American Academy of Arts and Sciences, and the American Philosophical Society.
Phase-separated Domains of mRNA Regulation and Epigenetic Memory
In the C. elegans germline, three distinct Argonaute (AGO) systems act in concert with nearly 1 million distinct guide RNAs to scan the entire transcriptome inclusive of the coding regions. These AGO systems include a scanning AGO (Piwi) and its genomically encoded piRNA cofactors, and two ‘memory’ AGO systems whose guides are synthesized and maintained by RNA-dependent RNA polymerase, (RdRP). The CSR-1 memory Argonaute targets but does not silence expressed germline mRNAs, while the WAGO memory Argonautes target mRNAs that are silenced in the germline. These RNA-surveillance pathways reside in, and help drive the association of perinuclear nuage, (P-granules), which form liquid-like condensates around nuclei in transcriptionally active germ cells.
In mature gametes, CSR-1 and WAGO small RNA complexes are transmitted in cytoplasmic P-granules (in oocytes), and in a perinuclear halo of Ago/RNPs in sperm. Interestingly piRNA targeting induces a muted recruitment of RdRP on CSR-1 targets, suggesting that CSR-1 protects its targets from silencing. In contrast, piRNAs induce the robust localized recruitment of RdRP on WAGO targets (or foreign mRNAs) to amplify WAGO guide RNAs that maintain (or induce) the silencing of the target mRNA. In addition to completely silencing its targets, experimental studies reveal that germline-expressed mRNAs can respond to artificially-increased piRNA targeting with partial reductions in mRNA and protein expression.
piRNAs are abundantly expressed in the germlines of diverse animals, and because they tolerate mismatched pairing, have a vast potential target space. Thus, the repertoire of piRNAs in metazoan germlines provides a wealth of potential post-transcriptional regulatory capacity. Our findings suggest that piRNAs can access and regulate mRNAs throughout the mature transcript, including the ORF, and that even the coding regions of mRNAs are free to sample regulatory inputs from piRNAs. Importantly, because piRNAs are expressed as independent genes, they are also free to evolve independently, unconstrained by the coding requirements of their target regions. Thus piRNAs may unlock a vast regulatory space, the coding regions of genes, for the control of gene expression. Thus all germline mRNAs appear to undergo Argonaute surveillance and regulation within peri-nuclear nuage as they export to the cytoplasm. The assembly of the nuage around the nuclear periphery may facilitate access of this scanning machinery to RNAs as they emerge, prior to Ribosome loading, thus permitting regulation through the entire transcript. The ubiquity of AGO scanning within nuage begs the question of whether the condensate forms to drive the function, or whether the function drives the form.
In other words, perhaps the massive scale of protein-RNA and RNA-RNA targeting within nuage drives the exclusion of water (which always happens on a molecular scale) but simply becomes evident in the light microscope—function drives the form?
Joseph (Jody) PUGLISI, PhD
Department of Structural Biology
Stanford University School of Medicine, USA
Member of the US National Academy of Sciences
Born and raised in New Jersey, Joseph (Jody) Puglisi received a B.A. degree in Chemistry in 1984 from The Johns Hopkins University and a Ph.D. in Biophysical Chemistry from UC Berkeley in 1989. After postdoctoral research in Strasbourg and MIT, he joined the faculty at UC Santa Cruz in Chemistry and Biochemistry in 1993. He moved to Stanford University in 1997 in the Department of Structural Biology.
Dr. Puglisi’s group investigates the role of RNA in cellular processes and disease. Their goal is to understand RNA function in terms of molecular structure and dynamics using a variety of biophysical and biological tools. They use nuclear magnetic resonance (NMR) spectroscopy to determine structures of biological molecules, and integrate structural understanding into further mechanistic and functional studies. They investigate dynamics using single-molecule approaches. Their goal is a unified picture of structure, dynamics and function. They are currently focused on the mechanism and regulation of translation, and the role of RNA in viral infections. A long-term goal is to target processes involving RNA with novel therapeutic strategies.
How mRNA controls the dynamics of translation
Translation by the ribosome is a central and final step in gene expression in all organisms. The goal of our research is to understand the underlying conformational and compositional dynamics of translation. We have applied single-molecule fluorescence and structural approaches to both prokaryotic and eukaryotic translation systems. We have explored the dynamics of translation initiation, elongation and termination. Here we will describe recent investigations of eukaryotic translation initiation in both yeast and humans that reveal the underlying dynamics of translational control.
Elisabetta Viani PUGLISI, PhD
Senior Research Scientist
Department of Structural Biology,
Stanford University School of Medicine, USA
Elisabetta Viani Puglisi was born in Parma and received her Laurea in Chemistry from the University of Parma in 1985, her Doctorate in Chemistry from the University of Parma in 1987 and Ph.D in Microbiology from the University of Brescia in 1994 for research performed at MIT. After postdoctoral research at UC Santa Cruz, she joined the Department of Structural Biology at Stanford in 1997. Her research focuses on the role of RNA structure in HIV replication.
Architecture of the HIV-1 reverse transcriptase initiation complex
Reverse transcription of the HIV-1 RNA genome into double-stranded DNA is a central step in infection and a common target of antiretroviral therapy. The reaction is catalyzed by viral reverse transcriptase (RT) that is packaged in an infectious virion along with 2 copies of dimeric viral genomic RNA and host tRNALys3, which acts as a primer for initiation of reverse transcription. Upon viral entry, initiation is slow and non-processive compared to elongation. We applied cryo-electron microscopy (cryo-EM) to determine the three-dimensional structure of the HIV RT initiation complex. RT is in an inactive polymerase conformation with open fingers and thumb and the nucleic acid primer-template complex is shifted away from the active site. The primer binding site (PBS) helix formed between tRNALys3 and HIV RNA lies in the cleft of RT, extended by additional pairing interactions. The 5’ end of the tRNA refolds and stacks on the PBS to create a long helical structure, while the remaining viral RNA forms two helical stems positioned above the RT active site, with a linker that connects these helices to the RNase H region of the PBS. Our results illustrate how RNA structure in the initiation complex alters RT conformation to decrease activity, highlighting a potential target for drug action.
Laura SEPP-LORENZINO, PhD
Vice President, Head of Nucleic Acid Therapies,
Vertex Pharmaceuticals, USA
Innovation at Vertex: Nucleic Acid Therapies as Transformative Medicines
Vertex is a global biotechnology company that invests in scientific innovation to create transformative medicines for people with serious and life-threatening diseases, through internal research and external partnerships. We discovered and developed the first medicines to treat the underlying cause of cystic fibrosis (CF), a rare, life-threatening genetic disease. In addition to cystic fibrosis, the following are diseases of further research interest: adrenoleukodystrophy, alpha-1 antitrypsin deficiency, polycystic kidney disease, and sickle cell disease.
Nucleic Acid Therapies expand our ability to target genes that are considered undruggable or difficult to drug with small molecules and biologics. Vertex established strategic research collaborations with Moderna and CRISPR to discover and develop potential new treatments aimed at the underlying genetic causes of human disease. Highlights from these efforts will be presented, in particular, our gene editing clinical program. As part of the CRISPR collaboration, we are co-developing CTX001, an investigational ex vivo CRISPR gene-edited therapy for patients suffering from β-thalassemia or sickle cell disease in which a patient’s hematopoietic stem cells are engineered to produce high levels of fetal hemoglobin (HbF; hemoglobin F) in red blood cells. HbF is a form of the oxygen carrying hemoglobin that is naturally present at birth and is then replaced by the adult form of hemoglobin. The elevation of HbF by CTX001 has the potential to alleviate transfusion-requirements for β-thalassemia patients and painful and debilitating sickle crises for sickle cell patients. CTX001 in on track to initiate a Phase 1/2 clinical study in SCD by the end of 2018 and are currently enrolling patients with transfusion dependent β-thalassemia in a Phase 1/2 trial in β-thalassemia in Europe.
Mikiko C. SIOMI, PhD
Professor, Department of Biological Sciences,
Graduate School of Science, The University of Tokyo, Japan
piRNA biogenesis and functions
Female sterile (1) Yb (Yb) is a primary component of Yb bodies, perinuclear foci known as the site for piRNA biogenesis in Drosophila ovarian somatic cells. Yb consists of Helicase C-terminal (Hel-C), RNA helicase and extended Tudor (eTud) domains. We previously showed that the RNA helicase domain is necessary for Yb RNA interaction, Yb body formation and piRNA biogenesis. Here, we investigated functions of two other domains and found that Hel-C and eTud are necessary for Yb to self-associate and to interact with RNAs and other Yb body components, respectively. Both domains were essential for Yb body formation and transposon silencing. Without eTud, piRNA production was totally impaired. Loss of Hel-C severely reduced the levels of transposon-targeting piRNAs while non-transposon-targeting piRNAs were fairly produced. Yb bodies seem to be liquid-like multivalent condensates whose assembly depends on Yb self-association and Yb-flam RNA binding.
Andreas STRASSER, PhD
Division Head, Molecular Genetics of Cancer,
The Walter and Eliza Hall Institute of Medical Research, Australia
Dr. Strasser is the head of the Molecular Genetics of Cancer Division at The Walter and Eliza Hall Institute (WEHI) in Melbourne, Australia. He completed his Master of Science and Ph.D. at the Basel Institute for Immunology and University of Basel in Switzerland before joining WEHI as a postdoctoral fellow in 1989. He studies programmed cell death and how defects in apoptosis cause cancer or autoimmune disease and impair the response of tumor cells to anti-cancer therapy. Dr. Strasser and his colleagues were the first to discover that abnormalities in cell death can cause cancer and autoimmune disease, including the discovery that BCL-2 collaborates with the MYC oncogene in tumorigenesis. Their studies have determined which pro-survival BCL-2 family member are essential for the sustained growth of which cancers. They also found that BCL-2 and death receptors regulate distinct pathways to apoptosis and have studied how the individual and overlapping roles of these two apoptotic pathways function in the immune system. Dr. Strasser discovered BIM and BMF, and was the first to show that BH3-only proteins are essential for the initiation of programmed cell death and stress-induced apoptosis. Based on this work, a collaboration between WEHI (including the Strasser group), Genentech, and AbbVie led to the development of the BCL-2 inhibitor Venetoclax/ABT-199, which is approved for treatment of refractory chronic lymphocytic leukemia, while a similar collaboration with Servier yielded the first potent and selective inhibitor of the cell death inhibitor MCL-1, currently in clinical trials.
Towards targeting MCL-1 for cancer therapy
Impaired apoptosis is considered one of the prerequisites for the development of most, if not all, cancers, but the mechanisms that guarantee the sustained survival of most cancer cells remain unknown. We examined the roles of individual pro-survival BCL-2 family members (when expressed under endogenous control) in lymphoma development. BCL-2 was found to be dispensable for the development of lymphoma. In contrast, loss of BCL-XL and even more remarkably, loss of a single allele of Mcl-1 greatly impaired lymphoma development in Eµ-Myc mice. Experiments with inducible knockout mice demonstrated that MCL-1 is essential for the sustained survival and expansion of MYC-driven malignant pore-B/B lymphoma. In collaboration with the pharma company Servier we have been testing S63845, a novel potent and selective inhibitor of MCL-1. S63845 was well tolerated in mice and could potently kill many human multiple myeloma, acute myeloid leukaemia and several other haematological and even solid cancer cell lines as single agent. These results reveal that the targeting of MCL-1 could be a promising strategy for the treatment of diverse cancers.
Contributors to this work and affiliations:
G Kelly*, S Glaser*, S Grabow*, JM Adams*, M Herold*, M Brennan*, O Geneste#, AL Maragno#, A Kotschy# and A Strasser*
*The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3050, Australia; #Servier, Paris, France
Bruce SULLENGER, PhD
Professor, Duke University Medical Center, USA
Dr. Sullenger’s research program focuses upon the discovery and development of therapeutic RNAs. His group has been a leader in the development of nucleic acid aptamers for inhibiting the activities of therapeutically important proteins including coagulation factors and platelet proteins. He is on the Board of Directors of the American Society of Gene and Cell Therapy and the Oligonucleotide Therapeutics Society and is Editor-in-Chief for the Society’s journal Nucleic Acid Therapeutics. Dr. Sullenger founded Regado Biosciences Inc, a company focused upon the development and commercialization of the aptamer-antidote technology his laboratory invented. The lead compound, an aptamer targeting factor IXa and its matched antidote, is in phase 3 clinical studies.
Dr. Sullenger earned his Bachelor of Science Degree from Indiana University (Bloomington, IN) and his Ph.D. from Cornell University Graduate School of Medical Sciences (New York, NY) working at the Memorial Sloan-Kettering Cancer Center. He performed postdoctoral studies at the University of Colorado (Boulder, CO) in the Department of Biochemistry in Dr. Thomas Cech’s laboratory.
Andrew YOO, PhD
Associate Professor, Department of Developmental Biology,
Washington University School of Medicine, USA
MicroRNAs as cell fate controllers and cellular reprogramming factors
Brain-enriched microRNAs (miRNAs), miR-9/9* and miR-124 (miR-9/9*-124), are potent cell fate regulators that can induce direct conversion (reprogramming) of non-neural human somatic cells to neurons. As such, ectopic expression of miR-9/9*-124 in human adult fibroblasts has been shown to directly target multiple components of chromatin modifiers leading to an extensive reconfiguration of the chromatin landscape while erasing the pre-existing fibroblast fate and adopting the neuronal identity. Our recent efforts in identifying direct targets of miR-9/9*-124 during neuronal conversion of human fibroblasts indicate multifactorial components in maintaining the non-neuronal fibroblast fate and repressors of neuronal fate. Some of the targeted pathways that converge to promote the fate switch include the downregulation of the USP14-EZH2-REST protein stability cascade, switching of subunit components of BAF chromatin remodeling complexes, alternative-splicing factors, and DNA methyltransferases. The resulting miRNA-induced chromatin then becomes permissive to the inputs of additional subtype-defining transcription factors that could guide the conversion to specific types of neurons including cortical neurons, striatal medium spiny neurons, and spinal cord motor neurons. Generation of specific types of human neurons through miRNA-mediated reprogramming then serves as an effective platform to model neurological disorders. Importantly for modeling late-onset diseases, directly converted human neurons have been shown to maintain the age signatures of fibroblast donors. As such, medium spiny neurons, a primary target of Huntington’s disease (HD), generated from direct conversion of HD fibroblasts reliably recapitulated hallmarks of adult-onset HD including the aggregation of Huntingtin protein, increased DNA damage, mitochondrial dysfunction and spontaneous neuronal death. Collectively, our studies demonstrate the potent activity of microRNAs as cell fate regulators and its utility to generate a patient-derived cellular platform to model diseases.