HNRNPH1

Data presented at International Society for Stem Cell Research conference shows unparalleled level of consistency across manufacturing of multiple human cell products World leading stem cell biologists call the technology “a watershed moment for biology” and “a true disruptive innovation in stem cell biology, much as CRISPR has been for genetics” bit.bio’s ioCells are already being used and enable large-scale experiments that underpin preclinical research and drug development The consistency of the cells is enabling scientific breakthroughs and applications ranging from biohybrid devices, novel targets for neurodegeneration, and cultured meat. CAMBRIDGE, England & BOSTON--(BUSINESS WIRE)-- bit.bio, a synthetic biology company focused on human cells, has achieved a milestone in the manufacture of human cells. Based on their opti-oxTM 1- based transcription factor reprogramming technology, bit.bio has reached a new level of precision and consistency of their induced pluripotent stem cells (iPSC)-derived cell products. The data that will be presented at the International Society for Stem Cell Research (ISSCR) conference show an unparalleled level of consistency with regards to multiple human cell products for research use. Samples of cells were selected from different manufacturing lots and time periods and different users carried out the opti-ox reprogramming step to manufacture the mature cell types. Analysis of the samples show fewer than 1% differentially expressed genes in bulk-mRNA sequencing experiments between manufacturing lots. This achievement sets a new standard for the manufacture of human cells. Commenting on the data, Dr Marius Wernig, Professor in the Departments of Pathology and Chemical and Systems Biology and Co-Director of the Institute for Stem Cell Biology and Regenerative Medicine at Stanford University and bit.bio SAB member said: “Out of 20,000 genes, not a single one is differentially expressed across three independently produced batches of bit.bio’s glutamatergic neurons. This seems like a watershed moment for biology. The team at bit.bio has achieved a level of consistency that is outside of the realm of what was thought possible. It is another proof point that transcription-factor based programming of cells can solve the challenges that have restricted the use of stem cell-derived cells in drug development and cell therapy.” Human stem cell pioneer Roger Pedersen, Senior Research Scientist at Stanford University School of Medicine and chair of bit.bio’s SAB said: “These new results are both compelling and biologically fascinating. bit.bio's opti-ox approach seems to remove interfering sources of stochasticity, thereby enabling deterministic reprogramming of iPSCs into somatic cells of any lineage. The homogeneity achieved by opti-ox is amazing, and I would never have thought such a high level of control over differentiation was possible. This is a true disruptive innovation in stem cell biology, much as CRISPR has been for genetics.” Bertie Göttgens, Professor and Director, University of Cambridge Stem Cell Institute commented: “bit.bio is an integral partner within the Cambridge Stem Cell community, where they have pushed the boundaries of what the field thought was possible with regards to consistent differentiation of human stem cells. From what I have seen, the data are astonishing. If this can be generalised to other cell types, these data will be seen as a landslide moment for the human stem cell field, which demonstrate that it is possible to industrialise the manufacture of human cells.” The consistency and scale of manufacturing achieved, with billions of cells now being produced on a daily basis, has opened doors to the creation of human cell standards. This has the potential to address the lack of reproducibility that has plagued the life sciences industry. By using reliable and standardised human cells, researchers worldwide can accelerate their studies and enhance the reliability of their findings. Thore Graepel, SVP of Computational biology at Altos Laboratories and bit.bio SAB member said: “I have never seen such consistency in biological data before. The data has made me rethink what level of consistency is possible in cell biology. Like in other application areas of machine learning, the quality of training data is of crucial importance and bit.bio’s cell products have the potential to super-charge AI models of biological systems.” Sir David Klenerman, Professor of Biochemistry University of Cambridge, 2020 Breakthrough Prize in Life Sciences recipient 2022 who is best known for his work on next-generation sequencing of DNA as the technological basis for Illumina, said: “This is a significant technological step forward, removing the cell-to-cell variation that has been a problem in the field.” Cells produced using the opti-ox technology have already been used in a range of research applications, including the identification of novel targets for neurodegenerative diseases such as Alzheimer’s,2,3. They have been used for the generation of biohybrid implant devices aiming to restore paralysed limb function4, and 3D printing of brain models5. The ability to produce cells at an industrial scale allows large-scale experimentation, such as high-throughput screening - for example, underpinning bit.bio’s partnership with leading contract research organisation Charles River Laboratories. This means scientists can now conduct experiments on a significantly larger scale in human models, leading to faster discoveries and a deeper understanding of cellular mechanisms. In addition, bit.bio is developing their technology for use in the field of cell therapy. Large scale manufacturing of human cells based on the ability to precisely reprogram iPSC into specific somatic cell types will enable off the shelf regenerative medicines that are available to millions of patients as mainstay treatments. Rahul Singhvi, CEO National Resilience stated: “This is an important breakthrough for the manufacture of human cells. The data are impressive and have set a new bar of what is possible. We are excited to support bit.bio in the development of their cell therapy pipeline.” Loïc Vincent, CSO Affini-T Therapeutics said: “A scalable source of consistent human cells will have a transformational impact on research, drug discovery, and cell therapy. It can tackle major challenges such as the reproducibility of scientific research all the way to scale up challenges for cell therapies and regenerative medicine. bit.bio's preclinical data show extraordinary consistency. The fact that this has been achieved across multiple cell types is also astonishing. bit.bio’s genomic safe harbour technology, opti-ox, is really pushing the boundaries.” Outside of medicine, the extraordinary scale enabled by opti-ox has enabled the manufacture of trillions of fat and muscle cells in a porcine background for cultured meat applications. Meatable held their first tasting in Singapore this year7 and is due to launch its first products by the end of the year. Meatable co-founder and CEO Krijn de Nood said: “Meatable is creating a world in which we can enjoy meat that is innocent. At the heart of our approach and enabling this dream is opti-ox. It allows us to manufacture at scale, and in the future reach price parity with meat derived from farmed animals. Whilst it is surprising to see these levels of consistency in the data, it is also what we experience on a daily basis. We found that opti-ox enables the manufacture of cells at all levels of scale, from the smallest experiment to the largest industrial bioreactor.” “The beauty of the data and how consistently bit.bio can produce human cells using our forward programming technology is mind-boggling” said Mark Kotter, CEO of bit.bio. “The team has developed solid processes and we haven’t had a single failed manufacturing run in the past two years. This is paving the way for groundbreaking research and future cell-based therapies. We are excited to collaborate with researchers and industry partners to unlock the full potential of human cells." Hermann Hauser KBE, Serial Entrepreneur and Venture Capitalist, e.g behind Acorn Computers, ARM, Solexa/Illumina and chair of the bit.bio Board said: “When I joined bit.bio I sensed that bit.bio’s opti-ox technology has the potential to create a category leader in the human cell space. The consistency that we detected in bit.bio’s products is astonishing. This is the bottom line if one gets everything right in a manufacturing process. I am proud of this fantastic achievement by the team.” bit.bio's human cell manufacturing technology is set to reshape the future of scientific research and biomedicine. By ensuring reliability, scalability, and reproducibility, bit.bio is driving forward the frontiers of cellular biology and transforming the possibilities of what can be achieved. Data for multiple cell types - glutamatergic neurons, sensory neurons and GABAergic neurons - will be presented at ISSCR, showing the approach may be generalisable to any human cell type. Notes to editors About the data This new data will be showcased in 4 posters and a talk at ISSCR TALK: Industrialising Cellular Reprogramming: Leveraging opti-ox™ Technology to Manufacture Human Cells with Unprecedented Consistency Marius Wernig and Mark Kotter Innovation Showcase - Thursday, 15 June 12:00 – 1:00pm, ET. ISSCR Transcription factor-mediated cellular reprogramming has emerged as a groundbreaking paradigm in developmental biology, challenging traditional theories and opening new avenues for further scientific innovations. Prof Wernig will discuss his pioneering work on cellular reprogramming, which opened up a new paradigm for cell identity, in which cellular states are driven by transcriptional events. His laboratory developed the first across germ layer reprogramming protocols highlighting the potential generalisability of this concept. His group demonstrated how transcription factor combinations are able to dictate cellular states and sub-cell identities. Based on this new paradigm, he developed new cellular models for translational research and outlined how reprogrammed cells could be used for therapeutic applications. Despite the benefits of cellular reprogramming, several challenges associated with conventional vector-based methods of transgene expression impact the efficiency, consistency and purity of the resulting cell populations, all of which need to be addressed for its potential to be truly realised in regenerative medicine. As Dr Kotter will outline, these challenges can be addressed by expressing reprogramming cassettes via genomic safe harbour (GSH) sites. GSH-mediated optimised inducible over-expression (opti-ox) enables highly controlled, consistent and scalable manufacturing of human iPSC-derived cells. Further, he will discuss how bit.bio has generalised this paradigm to generate cells from all three germ layers. The integration of opti-ox technology ensures increased precision and inducible control of transcription factor expression, setting a new standard for consistency in cell manufacturing. This breakthrough enables the manufacture of trillions of highly defined cells with unprecedented consistency offering the potential to transform the field of regenerative medicine and accelerate advancements in cell-based therapies. ○ POSTER: Generation Of Highly Pure, Consistent And Functional Inhibitory Gabaergic Neurons From Human Ipscs Using opti-ox Technology | Presenter Gianmarco Mastrogiovanni, Team leader ○ POSTER: Optimised and scalable reprogramming of human iPSCs to generate nociceptor sensory neurons for the study of pain mechanisms and neuropoathies | Presenter Dario Pacitti | Senior Scientist ○ POSTER: Modelling neurodegeneration using a human isogenic system: A next generation approach to study frontotemporal dementia and amyotrophic lateral sclerosis | Presenter Will Bernard​ | Director, Cell Type Development ○ POSTER: Rapid and consistent generation of functional microglia from reprogrammed hiPSCs to study mechanisms in neurodegeneration and neuroinflammation | Presenter Malathi Raman Srivastava​ | Snr Product Manager References: Matthias Pawlowski, Daniel Ortmann, Alessandro Bertero, Joana M. Tavares, Roger A. Pedersen, Ludovic Vallier, Mark R.N. Kotter 2017. Inducible and Deterministic Forward Programming of Human Pluripotent Stem Cells into Neurons, Skeletal Myocytes, and Oligodendrocytes, Stem Cell Reports, VOLUME 8, ISSUE 4, P803-812, APRIL 11, 2017. (16)X0005-7 Julie Qiaojin Lin, Deepak Khuperkar, Sofia Pavlou, Stanislaw Makarchuk, Nikolaos Patikas, Flora CY Lee, Julia M Zbiegly, Jianning Kang, Sarah F Field, David MD Bailey, Joshua L Freeman, Jernej Ule, Emmanouil Metzakopian, Marc-David Ruepp, Giovanna R Mallucci 2023. HNRNPH1 regulates the neuroprotective cold-shock protein RBM3 expression through poison exon exclusion, The EMBO Journal, 30 May 2023. Pavlou, S., Foskolou, S., Patikas, N. et al 2023. CRISPR-Cas9 genetic screen leads to the discovery of L-Moses, a KAT2B inhibitor that attenuates Tunicamycin-mediated neuronal cell death. Nature Scientific Reports 13, 3934 2023. Amy E. Rochford, Alejandro Carnicer-Lombarte, Malak Kawan, Amy Jin, Sam Hilton, Vincenzo F. Curto, Alexandra L. Rutz, Thomas Moreau, Mark R.N. Kotter, George G. Malliaras and Damiano G Barone 2023. Functional neurological restoration of amputated peripheral nerve using biohybrid regenerative bioelectronics, Science Advances, 22 Mar 2023, Vol 9, Issue 12, Zhou, L., Wolfes, A.C., Li, Y., Chan, D.C., Ko, H., Szele, F.G. and Bayley, H., 2020. Lipid‐bilayer‐supported 3D printing of human cerebral cortex cells reveals developmental interactions. Advanced Materials, 32(31), p.2002183. Press release, 11 May, 2023: Meatable holds its world-first tasting in Singapore with aim to launch in 2024. View source version on businesswire.com: Contacts April Six on behalf of bit.bio bitbio@aprilsix.com +44 7875 468942 Source: bit.bio View this news release online at:

Data from a new fetal brain atlas has helped a science team discover a collection of progenitor cells that give rise to aggressive 'group 3' medulloblastomas. Findings include identifying two genes that may serve as treatment targets. Mice with these tumors lived longer when bred to express reduced activity of these genes. Studying mice has filled encyclopedias with breakthrough medical discoveries. But when it comes to brain cancers, the little rodents have long exhibited fundamental limitations. Among them: the human cerebellum has 750 times as much surface area as a mouse, all of it laced with more types of progenitor cells that help the fetal brain grow during pregnancy. That means many things can go wrong in human brain development that simply cannot be seen by studying mice. Now a multinational team of scientists led by experts at Cincinnati Children's has developed an "atlas" of human fetal brain development so detailed that it details growth steps all the way down to changes occurring at the single-cell level. Details were published Nov. 30 in Nature. The investigators say this atlas will be a vital resource for brain research for years to come. In fact, it has already opened doors that someday may improve lives. "This study took the effort of 40 experts for nearly three years to complete. Their work included dozens of experiments using several of the very latest technologies in genomic science to reach this point," says senior author Qing Richard Lu, PhD, scientific director, Brain Tumor Center, Division of Experimental Hematology and Cancer Biology at Cincinnati Children's. "It was worth so much effort because this new map guided us to a targetable vulnerability for therapeutic intervention of aggressive medulloblastomas." Hunting cell-by-cell to pinpoint the birth of a killer Medulloblastomas are fast-growing malignant tumors that form in the back of the brain. They often disrupt balance and fine motor skills before going on to cause further damage. Overall, the five-year survival rate for this type of cancer is about 72%. However, about 27% of people with this form of cancer (most commonly children ages 4 to 16) develop "Group 3" aggressive medulloblastomas, which have five-year survival rates of about 20 to 30%. Now, thanks to data from the new fetal brain atlas, scientists have discovered a collection of progenitor cells (cells that make other types of cells) that give rise to group 3 medulloblastomas. This population of cells is rare in mouse brains, and exists only temporarily in the human brain as one type of cell morphs into another during fetal development. The team found these cells by comparing vast collections of data from brain tissue that developed tumors and tissue that did not. Now they have defined a hierarchy of cell formation as the brain grows, plus the trajectories or paths that progenitor cells follow as they form new types of brain cells or tumor cells under pathological conditions. Those transition moments appear critical. When healthy cell formation goes off-track, the fetal brain grows too many cells with excess activation of a potential cancer-causing gene called MYC. These cells wind up driving tumor formation. Cell analysis reveals possible way to slow tumor growth After identifying this new progenitor cell population as a potential root of group 3 medulloblastoma, the research team turned again to mouse models to learn if the newly discovered progenitor cells could be attacked. They found that eliminating or sharply reducing the activity of either of two genes, SOX11 or HNRNPH1, highly abundant in the progenitor cells, reversed tumor formation triggered by hyperactivity of the cancer-driving MYC gene. When mice were given human tumor cells with high levels of MYC but lacking these genes, tumor growth was inhibited and the animals survived longer. "This is exciting because these genes may serve as a potential target for future therapy of aggressive medulloblastoma," Lu says. Next steps Early success in mouse models represents the first step in a journey that could still take years to produce a treatment to help future children diagnosed with this aggressive form of brain tumor. Developing a test to detect the activity of the two MYC-regulating genes could help identify patients who may benefit from more aggressive chemotherapy in the clinic. Lu and colleagues also have begun work on identifying small-molecule compounds that can target the pathways supporting the aggressive subset of medulloblastoma. "While this study focuses on medulloblastoma, the new atlas will help accelerate understanding of other conditions that result from disruptions in healthy early brain development, such as autism, attention deficit-hyperactivity disorder (ADHD), and developmental dyslexia, and pediatric cerebellar damage," Lu says. This study was funded in part by grants from the Cincinnati Children's Research Foundation, the CancerFree Kids Foundation, the Pray-Hope-Believe Foundation, the TeamConnor Childhood Cancer Foundation, and the CureStartsNow Foundation.