Huode Bioengineering: Translating Innovative iPSC Technologies into Clinical-Grade Cell Therapies to Address Urgent Patient Needs

In November 2021, Hopstem Biotechnology announced the completion of its Series B financing round, amounting to hundreds of millions of yuan, led by Hillhouse Venture Capital and participated by Lilly Asia Ventures and YS Capital. To date, Hopstem Biotechnology has completed five rounds of financing, with investors including Longmen Capital, Hangjing Chuangke Venture Capital, Heli Investment, Cyberventure Investment, Data Capital, Ruisheng Investment, Huacheng Venture Capital, and YS Capital.

Among them, Longmen Capital has made multiple rounds of investment in Hopstem Biotechnology. Wang Haining, founding partner of Longmen Capital, stated, “Stem cells have long been an area of great interest to me, and I am optimistic about the potential clinical value of stem cell products. However, the domestic stem cell sector in China has long been plagued by disorder, compounded by regulatory challenges, with few teams genuinely committed to rigorous R&D and product development; consequently, we had previously refrained from investing. Induced pluripotent stem cells (iPSCs) present minimal ethical risks due to their origin and are more amenable to large-scale production, making them a highly promising pathway for the industrialization of stem cell therapies. With advances in iPSC technology and directed differentiation techniques, the treatment of refractory neurological diseases has become feasible.”

“Neural differentiation of iPSCs is highly challenging, and Hopstem Biotechnology is one of the few teams that have stably established a clinical-grade product portfolio.“Fan Bo is a leading young scientist in this field. What impresses me most is her deep understanding of and commitment to industrialization, making her one of the rare scientists who excel in both scientific research and industrial application. Due to our confidence in her team and strategic direction, we led a funding round last year and have further increased our investment this year.”

As Hopstem Biotechnology Completes a New Round of Financing, VCBeat Conducts an Exclusive Interview with Dr. Fan Jing, Founder of Hopstem Biotechnology

VCBeat:You began extensive research and application of pluripotent stem cell neural differentiation methods at Johns Hopkins University in the United States in 2012, and have 16 years of experience in studying neurodegenerative disease models and mechanisms of neurotoxicity. Why did you decide to found Hopstem Biotechnology in 2017?

Dr. Jing Fan:As the company’s founding member, I earned my bachelor’s degree from Peking University, obtained my Ph.D. in Neuroscience from the University of British Columbia (UBC) in Canada, and subsequently completed postdoctoral research at the Institute for Cell Engineering and the Department of Neurology at Johns Hopkins University in the United States. Over the past 16 years, my research has focused on elucidating the mechanisms and identifying therapeutic targets for neurodegenerative diseases and stroke. This extensive experience has not only deepened my understanding of these diseases but also prompted me to continually reflect on how to truly benefit patients. The pathophysiology of many neurological disorders is highly complex; despite years of dedicated efforts by numerous outstanding scientists worldwide, it remains challenging to fully unravel these mechanisms and pinpoint key molecular targets.

In 2016, our pioneering methods and research on neural differentiation of pluripotent stem cells, established at the Dawson Laboratory of Johns Hopkins University, were published in *Science Translational Medicine*. This approach has enabled my colleagues and me toUtilizing neural cells that closely mimic the composition and function of the human brain to investigate the mechanisms of stroke and neurodegenerative diseases and to screen potential therapeutics, this approach has also demonstrated significant promise for cell transplantation therapy in murine models of stroke.

During the same period, numerous companies were established in Japan and the United States and began developing iPSC-based cell products. At that time, I realized that,Direct transplantation of healthy functional cells to replace dead nerve cells in nerve injury and neurodegenerative diseases may represent a novel strategy.Such emerging technologies are likely toBypassing the black hole of molecular mechanisms to bring entirely new future therapeutic approaches to multiple disease areas.After six months of reflection and industry observation, I have made the firm decision to embark on entrepreneurship full-time, aiming to deliver effective treatments to patients as early as possible through iPSC and directed differentiation technologies.

Fortunately, in 2017, shortly after the company’s founding, Dr. Wang Anxin, who was engaged in iPSC-derived neural and glial differentiation research in Madison, Wisconsin, USA, decided to return to China. We quickly reached an agreement and jointly established Hopstem’s current patented technology and iPSC cell product development platform.

VCBeat:Global iPSC Cell Therapies Are in a Phase of Rapid Development: What Are the Market Opportunities and Pain Points?

Dr. Jing Fan:Currently, companies that have entered the clinical stage are primarily concentrated in Europe, the United States, and Japan. Existing clinical data have preliminarily demonstrated the safety and efficacy of iPSC-derived cell products, and major international pharmaceutical companies as well as leading capital firms are actively investing in this field.In China, companies developing iPSC-based cell therapies are in the preclinical research stage or have just begun industrialization.

iPSC reprogramming and directed differentiation technologies enable the mass production of functional human cells, holding broad application prospects across various disease areas.There is an urgent and substantial market demand for iPSC cell therapy, particularly in the fields of various central nervous system and ophthalmic diseases. For instance, in the stable hemiplegia phase after stroke, the stage of extensive dopaminergic neuron loss in Parkinson’s disease, and non-hereditary degenerative eye diseases, the advantages of iPSC cell therapy are unattainable by small-molecule drugs, biologics, or gene therapies.

Furthermore, in the field of tumor immunotherapy, gene editing and clonal screening of iPSC cell lines can be employed to achieve mass production and quality control of off-the-shelf CAR-T, CAR-NK, and CAR-macrophage cell products with enhanced cytotoxicity, greater stability and consistency, and improved safety profiles. This approach significantly reduces the cost of such cell therapies, thereby benefiting all patients.

Currently, the key pain points in the iPSC cell therapy industry lie in the scarcity of GMP-compliant materials, automated equipment, third-party testing services, and specialized industry talent, as the sector is still in its early stages. Of course, this is a common challenge for emerging industries. With rapid development and sustained investment, these issues are being gradually resolved, while corresponding regulatory frameworks and quality standards continue to be refined.

VCBeat:iPSC Cell Therapy: High Barriers and Complex Processes—What Are the Specific R&D Hurdles, and How Does Hopstem Biotechnology Overcome Them?

Dr. Jing Fan:One of the biggest R&D barriers isGMP-compliant iPSC cell lines and reprogramming patents.Patents for iPSC reprogramming are predominantly held by institutions such as the Whitehead Institute, Cellular Dynamics International, Factor Bioscience, and RIKEN. As companies approach the commercialization stage, they will likely need to secure licenses for the iPSC reprogramming patents utilized in early-stage product development, with associated costs and uncertainties posing significant risks.

After being alerted by investors in the early stages, Hopstem Biotechnology engaged in negotiations with multiple companies, including Japan’s iPSC Academy, regarding commercial licensing of cell lines and patents. Ultimately, in 2019, through independent R&D and freedom-to-operate (FTO) analysis, it successfully established its ownmRNA and small molecule combination-based iPSC reprogramming method, for which a series of patents have been filed.Among them, the application in Australia has been granted, while the application in China has been published and entered the national phase via the PCT.

This means that our clinical products can achieve global development without being constrained by external parties, and the foreign GMP-compliant iPSC cell lines established through our proprietary patents can be used for licensing and collaboration both domestically and internationally. The company also benefited from the INTERACT meeting with the U.S. FDA in 2019, establishing guidelines based on communication feedback and existing domestic regulatory principles.Clinical-Grade iPSC Cell Lines and Cell Banks。

Secondly,CMC Development and Quality Standard Establishment for iPSC-Derived Cell Products Also Face High Barriers. For example, after extensive testing of numerous clinical-grade reagents and processes, the team ultimately stabilized a manufacturing process capable of producing high-quality, large-scale neural cell products. Furthermore, iPSC-derived cell products exhibit many characteristics distinct from primary cell products, and there are currently no corresponding quality control methods or quality standards in place—for instance, for detecting residual iPSCs in the final product, characterizing cellular composition, and performing whole-genome mutation detection and validation. It took us nearly three years of extensive QC method development work to gradually establish a comprehensive suite of assay methods.

The transition from research and technology to industry and clinical application is a challenging yet highly rewarding transformation. It requires the team to possess rapid learning capabilities, undergo a painful shift in mindset, and, most importantly, maintain an unwavering commitment to delivering stable, consistent, safe, and effective cell-based therapies for patients.

VCBeat:What are the advantages of Hopstem Biotechnology’s second-generation technology for inducing differentiation of pluripotent stem cells, and which limitations of conventional technologies can it address?

Dr. Jing Fan:Hopstem Biotechnology’s second-generation RONA neuronal differentiation method is an optimized approach derived from the first-generation method published in *Science Translational Medicine* in 2016, achieved through the screening of small molecules that promote neuronal differentiation and maturation.

First, second-generation methods still possess advantages not found in conventional EB-based methods, monolayer cultures, or direct transdifferentiation approaches. These include the ability to authentically recapitulate the cellular composition and developmental processes of the brain, yielding balanced neuronal populations capable of long-term survival both in vitro and in vivo (for over three years). Furthermore, these cells exhibit a degree of functional maturity closely resembling that of the adult human brain and can form stable neural networks.

Traditional differentiation methods generally yield only pure excitatory or inhibitory forebrain neurons, which lack mutual facilitation and regulatory functions. These cells are insufficiently mature and stable, potentially hindering the formation of neural network connections upon transplantation and even posing a risk of epilepsy. Furthermore, traditional neural differentiation methods typically require more than 30 days of NPC differentiation to achieve relatively mature electrophysiological properties, whereasThe second-generation method can shorten the differentiation and maturation time of neural cells to approximately 7–14 days, resulting in synchronized electrical firing (formation of neural networks). Furthermore, these cells can differentiate into various functional neuron types in vivo and gradually integrate into the neural networks of stroke animal models.

It should be emphasized that although the RONA differentiation method is relatively complex, in addition toIn addition to being the only method currently available for obtaining cells with a normal forebrain composition and a high degree of functional maturity, it also achieves higher purity and safety due to its unique intermediate step of physically purifying the neuroectoderm (for which equipment is already available to support a closed, fully automated process).

Hopstem Biotechnology can produce dopaminergic neurons with over 98% Foxa2 positivity, which further differentiate into cells with over 95% positivity for both TH and DAT (as verified by flow cytometry and single-cell transcriptomics). This contrasts with traditional differentiation methods, which typically yield dopaminergic cells with only 30% purity, necessitating additional purification steps such as magnetic bead sorting. Furthermore, compared to traditional methods, the second-generation method yields dopaminergic neurons with higher viability and functionality, thereby facilitating better post-transplant survival and functional integration.

VCBeat:How does Hopstem Biotechnology approach its strategy for disease area selection and R&D pipeline layout? What is the current progress of its pipeline development?

Dr. Jing Fan:Stroke, hemiplegia in the stable phase following traumatic brain injury, Parkinson’s disease (PD), Alzheimer’s disease (AD), and amyotrophic lateral sclerosis (ALS) represent significant and urgent unmet clinical needs, as well as areas where cell replacement therapy can play a substantial role. With extensive R&D experience and robust technological reserves in neurological disorders, Hopstem Biotechnology has focused its current pipeline primarily on diseases of the nervous system.

The Company’s first product for the treatment of ischemic stroke, hNPC01, is undergoing preclinical safety and efficacy studies, with Investigational New Drug (IND) applications in China and the United States expected to be filed in 2022. Subsequent pipeline candidates are in the Chemistry, Manufacturing, and Controls (CMC) development stage. Our developedThe first forebrain neural progenitor cell product for stroke is expected to be the most advanced and fastest-acting globally, while the dopamine precursor cells for Parkinson’s disease also offer superior quality advantages over previous products.

The human iPSC-derived cell product for the treatment of ocular diseases, jointly developed by the Company and Nuofo Biotechnology Co., Ltd., is also in the CMC development stage. Although the Company’s CMC platform can rapidly translate other categories of iPSC cell products, we will carefully select and gradually expand our product pipeline and collaborations based on thorough evaluations and the Company’s growth trajectory, ensuring that our lead products reach patients as soon as possible.

Moving forward, the company’s short-term objective is to advance its existing products and pipeline into clinical stages. Its long-term strategic plan is to continually enrich its product pipeline and expand into international markets, driving continuous innovation to provide safe, effective, and affordable iPSC-derived cell therapies for patients worldwide.