Institute of Medical Science

Xenetic to supply recombinant DNase I to Tokyo Medical University for evaluation as a treatment of Ewing sarcoma in unique preclinical model FRAMINGHAM, MA / ACCESSWIRE / October 17, 2024 / Xenetic Biosciences, Inc. (NASDAQ:XBIO) ("Xenetic" or the "Company"), a biopharmaceutical company focused on advancing innovative immuno-oncology technologies addressing hard to treat oncology indications, today announced it has entered into a Materials Transfer Agreement with Tokyo Medical University to advance the development of its systemic DNase program. Under the terms of the agreement, Professor Takuro Nakamura of the Department of Experimental Pathology, Institute of Medical Science at Tokyo Medical University will lead the research program evaluating the effects of human recombinant DNase I (rhDNase I) when given in combination with chemotherapy in a proprietary immunocompetent preclinical mouse model of Ewing sarcoma. Professor Takuro Nakamura's proprietary immunocompetent Ewing sarcoma model encompasses the biological characteristics, morphology and gene expression profiles of human Ewing sarcoma and has demonstrated translational relevance. Ewing sarcoma is an aggressive orphan pediatric cancer that grows in bones or soft tissues, accounting for between 2 to 3 percent of all childhood cancers. There is a lack of effective treatment options for children with recurrent and metastatic disease where the five-year survival rate is only 20 to 30 percent for patients that have relapsed. Clinical studies conducted at Tel Aviv Medical Center between 2010 and 2021 [1] showed that the formation of neutrophil extracellular traps (NETs) in the tumor microenvironment of Ewing sarcoma is an independent prognostic factor, with a clear association between NETs burden and poor prognosis. According to research from these clinical studies, elevated levels of NETs at diagnosis predicted a poor response to neoadjuvant chemotherapy, relapse, and death from the disease. Xenetic's proprietary recombinant DNase I is an enzyme that digests NETs in tumor microenvironment. The preclinical studies are designed to evaluate the efficacy of DNase to reduce NETs burden and to increase the efficacy of chemotherapy given in an adjuvant setting. James Parslow, Interim Chief Executive Officer and Chief Financial Officer of the Company stated, "As part of our overall development strategy, we aim to leverage relationships like the one established with Tokyo Medical University. Our commitment to the DNase program remains steadfast, and we are pleased to enter into this agreement to further expand our growing body of data." About DNase-Based Oncology Platform Xenetic's DNase-based oncology platform is designed to target NETs, which are weblike structures composed of extracellular chromatin coated with histones and other proteins. In cancer, NETs are expelled by activated neutrophils into the tumor microenvironment and blood, thereby promoting cancer spread and local and systemic immunosuppression. Reduction of NETs burden via application of Xenetic's proprietary recombinant human DNase I has been shown to improve efficacy of immunotherapy, adoptive cell therapy and chemotherapy in preclinical animal models. About Xenetic Biosciences Xenetic Biosciences, Inc. is a biopharmaceutical company focused on advancing innovative immune-oncology technologies addressing hard to treat cancers. The Company's DNase platform is designed to improve outcomes of existing treatments, including immunotherapies, by targeting neutrophil extracellular traps (NETs), which are involved in cancer progression. Xenetic is currently focused on advancing its systemic DNase program into the clinic as an adjunctive therapy for pancreatic carcinoma and locally advanced or metastatic solid tumors. For more information, please visit the Company's website at www.xeneticbio.com and connect on X , LinkedIn , and Facebook . Forward-Looking Statements This press release contains forward-looking statements that we intend to be subject to the safe harbor provisions of the Private Securities Litigation Reform Act of 1995. All statements contained in this press release other than statements of historical facts may constitute forward-looking statements within the meaning of the federal securities laws. These statements can be identified by words such as "expects," "plans," "projects," "will," "may," "anticipates," "believes," "should," "intends," "estimates," "remain," "focus", and other words of similar meaning, including, but not limited to, all statements regarding the Materials Transfer Agreement with Tokyo Medical University, including the design of preclinical studies, our aim to leverage relationships like the one established with Tokyo Medical University, our commitment to the DNase program, and our expectations that such agreement will further expand our growing body of data, and all statements regarding expectations for our DNase-base oncology platform, including statements regarding our focus on advancing innovative immune-oncology technologies addressing hard to treat cancers, the DNase platform improving outcomes of existing treatments, NETs, and our focus on advancing our systemic DNase program into the clinic as an adjunctive therapy for pancreatic carcinoma and locally advanced or metastatic solid tumors. Any forward-looking statements contained herein are based on current expectations and are subject to a number of risks and uncertainties. Many factors could cause our actual activities, performance, achievements, or results to differ materially from the activities and results anticipated in forward-looking statements. Important factors that could cause actual activities, performance, achievements, or results to differ materially from such plans, estimates or expectations include, among others, (1) failure to achieve the expected outcomes and realize the anticipated benefits from our collaboration agreements, including the agreement with Tokyo Medical University; (2) unexpected costs, charges or expenses resulting from our manufacturing and collaboration agreements, including the agreement with Tokyo Medical University; (3) unexpected costs, charges or expenses resulting from the licensing of the DNase platform; (4) uncertainty of the expected financial performance of the Company following the licensing of the DNase platform; (5) failure to realize the anticipated potential of the DNase or PolyXen technologies; (6) the ability of the Company to obtain funding and implement its business strategy; and (7) other risk factors as detailed from time to time in the Company's reports filed with the SEC, including its annual report on Form 10-K, periodic quarterly reports on Form 10-Q, current reports on Form 8-K and other documents filed with the SEC. The foregoing list of important factors is not exclusive. In addition, forward-looking statements may also be adversely affected by general market factors, general economic and business conditions, including potential adverse effects of public health issues, such as the COVID-19 outbreak, and geopolitical events, such as the Russian invasion of Ukraine and conflict in the Middle East, on economic activity, competitive product development, product availability, federal and state regulations and legislation, the regulatory process for new product candidates and indications, manufacturing issues that may arise, patent positions, litigation, and shareholder activism, among other factors. The forward-looking statements contained in this press release speak only as of the date the statements were made, and the Company does not undertake any obligation to update forward-looking statements, except as required by law. Contact: JTC Team, LLC Jenene Thomas (908) 824-0775 xbio@jtcir.com 1 https://onlinelibrary.wiley.com/doi/full/10.1111/cas.15992 SOURCE: Xenetic Biosciences, Inc. Released October 17, 2024

A suite of three innovations enables high-resolution, high-throughput imaging of human brain tissue at a full range of scales and mapping connectivity of neurons at single cell resolution. To demonstrate the advance, researchers compared a brain region in an Alzheimer's and a control sample. Observing anything and everything within the human brain, no matter how large or small while it is fully intact, has been an out-of-reach dream of neuroscience for decades, but in a new study in Science, an MIT-based team describes a technology pipeline that enabled them to finely process, richly label and sharply image full hemispheres of the brains of two donors -- one with Alzheimer's and one without -- at high resolution and speed. "We performed holistic imaging of human brain tissues at multiple resolutions from single synapses to whole brain hemispheres and we have made that data available," said senior and corresponding author Kwanghun Chung, associate professor in The Picower Institute for Learning and Memory, the Departments of Chemical Engineering and Brain and Cognitive Sciences, and the Institute for Medical Engineering and Science at MIT. "This technology pipeline really enables us to analyze the human brain at multiple scales. Potentially this pipeline can be used for fully mapping human brains." The new study does not already present a comprehensive map or atlas of the entire brain, in which every cell, circuit and protein is identified and analyzed, but with full hemispheric imaging, it demonstrates an integrated suite of three technologies to enable that and other long-sought neuroscience investigations. The research provides a "proof of concept" by showing numerous examples of what the pipeline makes possible, including sweeping landscapes of thousands of neurons within whole brain regions, diverse forests of cells each in individual detail, and tufts of subcellular structures nestled among extracellular molecules. The researchers also present a rich variety of quantitative analytical comparisons focused on a chosen region within the Alzheimer's and non-Alzheimer's hemispheres. The importance of being able to image whole hemispheres of human brains intact and down to the resolution of individual synapses (the teeny connections that neurons forge to make circuits) is two-fold for understanding the human brain in health and disease, Chung said. On one hand, it will enable scientists to conduct integrated explorations of questions using the same brain, rather than having to, for example, observe different phenomena in different brains, which can vary significantly, and then trying to construct a composite picture of the whole system. A key feature of the new technology pipeline is that analysis doesn't degrade the tissue. On the contrary, it makes the tissues extremely durable and repeatedly re-labelable to highlight different cells or molecules as needed for new studies for potentially years on end. In the paper Chung's team demonstrates using 20 different antibody labels to highlight different cells and proteins but they are already expanding that to a hundred or more. "We need to be able to see all these different functional components -- cells, their morphology and their connectivity, subcellular architectures, and their individual synaptic connections -- ideally within the same brain, considering the high individual variabilities in the human brain and considering the precious nature of human brain samples," Chung said. "This technology pipeline really enables us to extract all these important features from the same brain in a fully integrated manner." On the other hand, the pipeline's relatively high scalability and throughput (imaging a whole brain hemisphere once it is prepared takes 100 hours rather than many months) means that it is possible to create many samples to represent different sexes, ages, disease states and other factors that can enable robust comparisons with increased statistical power. Chung said he envisions creating a brain bank of fully imaged brains that researchers could analyze and re-label as needed for new studies to make more of the kinds of comparisons he and co-authors made with the Alzheimer's and non-Alzheimer's hemispheres in the new paper. Three key innovations Chung said the biggest challenge he faced in achieving the advances described in the paper was building a team at MIT that included three especially talented young scientists, each a co-lead author of the paper because of their key roles in producing the three major innovations. Ji Wang, a mechanical engineer and former postdoc, developed the "Megatome," a device for slicing intact human brain hemispheres so finely that there is no damage to it. Juhyuk Park, a materials engineer and former postdoc, developed the chemistry that makes each brain slice clear, flexible, durable, expandable, and quickly, evenly and repeatedly labelable -- a technology called "mELAST." Webster Guan, a former MIT chemical engineering graduate student with a knack for software development, created a computational system called "UNSLICE" that can seamlessly reunify the slabs to reconstruct each hemisphere in full 3D down to the precise alignment of individual blood vessels and neural axons (the long strands they extend to forge connections with other neurons). No technology allows for imaging whole human brain anatomy at subcellular resolution without first slicing it because it is very thick (it's 3,000 times the volume of a mouse brain) and opaque. But in the Megatome, tissue remains undamaged because Wang, who is now at a company Chung founded called LifeCanvas Technologies, engineered its blade to vibrate side to side faster and yet sweep wider than previous vibratome slicers. Meanwhile she also crafted the instrument to stay perfectly within its plane, Chung said. The result are slices that don't lose anatomical information at their separation or anywhere else. And because the vibratome cuts relatively quickly and can cut thicker (and therefore fewer) slabs of tissue, a whole hemisphere can be sliced in a day, rather than months. A major reason why slabs in the pipeline can be thicker comes from mELAST. Park engineered the hydrogel that infuses the brain sample to make it optically clear, virtually indestructible and compressible and expandable. Combined with other chemical engineering technologies developed in recent years in Chung's lab, the samples can then be evenly and quickly infused with the antibody labels that highlight cells and proteins of interest. Using a light sheet microscope the lab customized, a whole hemisphere can be imaged down to individual synapses in about 100 hours, the authors report in the study. Park is now an assistant professor at Seoul National University in South Korea. "This advanced polymeric network, which fine-tunes the physicochemical properties of tissues, enabled multiplexed multiscale imaging of the intact human brains," Park said. After each slab has been imaged, the task is then to restore an intact picture of the whole hemisphere computationally. Guan's UNSLICE does this at multiple scales. For instance, at the middle, or "meso" scale, it algorithmically traces blood vessels coming into one layer from adjacent layers and matches them. But it also takes an even finer approach. To further register the slabs, the team purposely labeled neighboring neural axons in different colors (like the wires in an electrical fixture). That enabled UNSLICE to match layers up based on tracing the axons, Chung said. Guan is also now at LifeCanvas. In the study the researchers present a litany of examples of what the pipeline can do. The very first figure demonstrates that the imaging allows one to richly label a whole hemisphere and then zoom in from the wide scale of brainwide structures to the level of circuits, then individual cells and then subcellular components such as synapses. Other images and videos demonstrate how diverse the labeling can be, revealing long axonal connections and the abundance and shape of different cell types including not only neurons but also astrocytes and microglia. Exploring Alzheimer's For years Chung has collaborated with co-author Matthew Frosch, an Alzheimer's researcher and director of the brain bank at Massachusetts General Hospital, to image and understand Alzheimer's disease brains. With the new pipeline established they began an open-ended exploration, first noticing where within a slab of tissue they saw the greatest loss of neurons in the disease sample compared to the control. From there, they followed their curiosity -- as the technology allowed them to do -- ultimately producing a series of detailed investigations described in the paper. "We didn't lay out all these experiments in advance," Chung said. "We just started by saying, 'OK, let's image this slab and see what we see.' We identified brain regions with substantial neuronal loss so let's see what's happening there. 'Let's dive deeper.' So we used many different markers to characterize and see the relationships between pathogenic factors and different cell types. "This pipeline allows us to have almost unlimited access to the tissue," Chung said. "We can always go back and look at something new." They focused most of their analysis in the orbitofrontal cortex within each hemisphere. One of the many observations they made was that synapse loss was concentrated in areas where there was direct overlap with amyloid plaques. Outside of areas of plaques the synapse density was as high in the brain with Alzheimer's as in the one without the disease. With just two samples, Chung said, the team is not offering any conclusions about the nature of Alzheimer's disease, of course, but the point of the study is that the capability now exists to fully image and deeply analyze whole human brain hemispheres to enable exactly that kind of research. Notably, the technology applies equally well to many other tissues in the body, not just brains. "We envision that this scalable technology platform will advance our understanding of the human organ functions and disease mechanisms to spur development of new therapies," the authors conclude. In addition to Park, Wang, Guan, Chung and Frosch, the paper's other authors are Lars A. Gjesteby, Dylan Pollack, Lee Kamentsky, Nicholas B. Evans, Jeff Stirman, Xinyi Gu, Chuanxi Zhao, Slayton Marx, Minyoung E. Kim, Seo Woo Choi, Michael Snyder, David Chavez, Clover Su-Arcaro, Yuxuan Tian, Chang Sin Park, Qiangge Zhang, Dae Hee Yun, Mira Moukheiber, Guoping Feng, X. William Yang, C. Dirk Keene, Patrick R. Hof, Satrajit S. Ghosh, and Laura J. Brattain. The main funding for the work came from the National Institutes of Health, The Picower Institute for Learning and Memory, The JPB Foundation, and the NCSOFT Cultural Foundation.

A recent study in genetic control elements revealed around 11,000 gene regulators active in every cell type, also known as housekeeping cis-regulatory elements (HK-CREs). These elements are vital in maintaining cellular stability beyond conventional gene regulation, influencing diverse cellular functions across healthy cell types. Moreover, a subset of these housekeeping elements, particularly those related to zinc finger genes, was found to have reduced activity in diverse cancers, suggesting their role as potential housekeeping tumor suppressors. Technological advancements have enabled scientists to comprehensively explore genetic control elements, unraveling the complexities of gene activation mechanisms in our genetic code. New evidence challenges the simplistic view that cis-regulatory elements (CREs) are mere on/off switches for genes, emphasizing their ability to exhibit complex behaviors, such as the simultaneous enhancement of gene activity and initiation of gene transcription, e.g., simultaneous enhancer and promoter activities. These switches aren't only important for the enhancement of specific genes but are crucial for the basic functions that keep our cells healthy. Now, a study conducted in Japan has revealed the existence of around 11,000 vital genetic switches active in every cell type -- housekeeping cis-regulatory elements (HK-CREs) -- that play a role in maintaining the stability and function of our cells, far beyond the regulation of housekeeping genes. This study was performed by scientists from the Laboratory of Functional Analysis in silico (Nakai-lab) at The Institute of Medical Science, The University of Tokyo, Japan: Professor Kenta Nakai, head of the laboratory, and Dr. Martin Loza, Assistant Professor, in collaboration with Dr. Alexis Vandenbon, Associate Professor, from the Institute of Life and Medical Sciences, Kyoto University, Japan. Their work was published in Nucleic Acids Research on December 12, 2023. Discussing his motivation behind this study, Dr. Loza states, "Given the significant association between cancer and mutations in epigenetic components, every small insight we gain could be key in the ongoing battle against this disease, which has tragically claimed innumerable lives. Through extensive bioinformatics analyses, we aimed to emphasize HK-CREs profound impact on fundamental cellular processes, including their potential as essential housekeeping tumor suppressors." The research team found that HK-CREs were not solely confined to regulating the well-studied housekeeping genes (HKGs), which only constituted less than 20% of the genes associated with these elements. Instead, these elements predominantly resided within core promoter regions of many more genes (around 8,000), indicating a broader regulatory role beyond typical housekeeping gene functions. By employing bioinformatics analyses and levering diverse public datasets, the team validated the robustness of HK-CREs across 50 randomly selected healthy cell types, confirming the location of HK-CREs within the genome. These elements were highly conserved, residing in unmethylated CpG-rich regions, a trait strongly associated with their housekeeping regulatory function. Sharing his concerns about the analysis, Dr. Lozastates, "By leveraging bioinformatics analyses of multiomics data, we offer an approach to harnessing publicly available datasets for exploring diverse biological mechanisms. We anticipate that employing similar workflows can significantly streamline analyses, cutting back both time and financial investments needed for comprehensive studies involving new data." The team remarked on the intricate cooperative interactions among housekeeping core promoters (HK-CPs), forming complex regulatory networks through promoter-promoter interactions. These observations hint at the significant influence of such interactions not only on HKGs but also on genes specific to various cell types. Turning their attention to cancer cells, researchers discovered a subset of HK-CREs displaying reduced activity in diverse cancer subtypes due to aberrant methylation, particularly those linked to zinc finger genes clustered in sub-telomere regions of chromosome 19. Identifying genes such as ZNF135, ZNF154, ZNF667, and ZNF667-AS1 under the influence of these foundational core promoters, the research suggests their potential as housekeeping tumor suppressor genes. "Genes detected in our study have exhibited decreased activity in multiple cancer cell lines, and survival analysis across various cancer projects have revealed significant increases in survival probability in diverse cancer types like pancreas adenocarcinoma and uveal melanoma," states Dr. Loza. In essence, the results of this research have uncovered a previously unknown class of HK-CREs critical for cellular stability, extending their influence beyond housekeeping gene regulation. "Our discovery on housekeeping tumor suppressor genes unveils a novel avenue in cancer therapy, harnessing the intrinsic elements within the DNA of every cell. Future approaches to cancer treatment, focusing on these housekeeping tumor suppressor genes, offer a unique solution that could potentially target a broad range of cancers, sidestepping the challenges associated with personalized medicine," remarks Dr. Loza. He further adds, "Our findings on housekeeping cis-regulatory elements fill a big gap in the current knowledge regarding gene regulatory processes. We anticipate that our findings will enhance the understanding of these processes and serve as a valuable resource for researchers striving to uncover elements inherent in the genome for combating various diseases." Technological advancements have enabled scientists to comprehensively explore genetic control elements, unraveling the complexities of gene activation mechanisms in our genetic code. New evidence challenges the simplistic view that cis-regulatory elements (CREs) are mere on/off switches for genes, emphasizing their ability to exhibit complex behaviors, such as the simultaneous enhancement of gene activity and initiation of gene transcription, e.g., simultaneous enhancer and promoter activities. These switches aren't only important for the enhancement of specific genes but are crucial for the basic functions that keep our cells healthy. Now, a study conducted in Japan has revealed the existence of around 11,000 vital genetic switches active in every cell type -- housekeeping cis-regulatory elements (HK-CREs) -- that play a role in maintaining the stability and function of our cells, far beyond the regulation of housekeeping genes. This study was performed by scientists from the Laboratory of Functional Analysis in silico (Nakai-lab) at The Institute of Medical Science, The University of Tokyo, Japan: Professor Kenta Nakai, head of the laboratory, and Dr. Martin Loza, Assistant Professor, in collaboration with Dr. Alexis Vandenbon, Associate Professor, from the Institute of Life and Medical Sciences, Kyoto University, Japan. Their work was published in Nucleic Acids Research on December 12, 2023. Discussing his motivation behind this study, Dr. Loza states, "Given the significant association between cancer and mutations in epigenetic components, every small insight we gain could be key in the ongoing battle against this disease, which has tragically claimed innumerable lives. Through extensive bioinformatics analyses, we aimed to emphasize HK-CREs profound impact on fundamental cellular processes, including their potential as essential housekeeping tumor suppressors." The research team found that HK-CREs were not solely confined to regulating the well-studied housekeeping genes (HKGs), which only constituted less than 20% of the genes associated with these elements. Instead, these elements predominantly resided within core promoter regions of many more genes (around 8,000), indicating a broader regulatory role beyond typical housekeeping gene functions. By employing bioinformatics analyses and levering diverse public datasets, the team validated the robustness of HK-CREs across 50 randomly selected healthy cell types, confirming the location of HK-CREs within the genome. These elements were highly conserved, residing in unmethylated CpG-rich regions, a trait strongly associated with their housekeeping regulatory function. Sharing his concerns about the analysis, Dr. Lozastates, "By leveraging bioinformatics analyses of multiomics data, we offer an approach to harnessing publicly available datasets for exploring diverse biological mechanisms. We anticipate that employing similar workflows can significantly streamline analyses, cutting back both time and financial investments needed for comprehensive studies involving new data." The team remarked on the intricate cooperative interactions among housekeeping core promoters (HK-CPs), forming complex regulatory networks through promoter-promoter interactions. These observations hint at the significant influence of such interactions not only on HKGs but also on genes specific to various cell types. Turning their attention to cancer cells, researchers discovered a subset of HK-CREs displaying reduced activity in diverse cancer subtypes due to aberrant methylation, particularly those linked to zinc finger genes clustered in sub-telomere regions of chromosome 19. Identifying genes such as ZNF135, ZNF154, ZNF667, and ZNF667-AS1 under the influence of these foundational core promoters, the research suggests their potential as housekeeping tumor suppressor genes. "Genes detected in our study have exhibited decreased activity in multiple cancer cell lines, and survival analysis across various cancer projects have revealed significant increases in survival probability in diverse cancer types like pancreas adenocarcinoma and uveal melanoma," states Dr. Loza. In essence, the results of this research have uncovered a previously unknown class of HK-CREs critical for cellular stability, extending their influence beyond housekeeping gene regulation. "Our discovery on housekeeping tumor suppressor genes unveils a novel avenue in cancer therapy, harnessing the intrinsic elements within the DNA of every cell. Future approaches to cancer treatment, focusing on these housekeeping tumor suppressor genes, offer a unique solution that could potentially target a broad range of cancers, sidestepping the challenges associated with personalized medicine," remarks Dr. Loza. He further adds, "Our findings on housekeeping cis-regulatory elements fill a big gap in the current knowledge regarding gene regulatory processes. We anticipate that our findings will enhance the understanding of these processes and serve as a valuable resource for researchers striving to uncover elements inherent in the genome for combating various diseases."