Improved Delivery and Function of Myoblasts via Soluble TIPS Microcarriers

Lead Research Organisation: University College London
Department Name: Medicine

Abstract

Incontinence is a debilitating condition with devastating social, economic and psychological consequences. A common cause is damage sustained during childbirth to the muscles that normally provide control of bowel movements. Current medical and surgical treatments are often inadequate. Emerging treatments involving the use of a patient's own muscle cells offer the possibility of restoring function to the damaged muscle, but improved methods for delivering the cells to ensure they survive and repair tissue are needed.

The aim of the proposed research is to demonstrate, through proof of principle, that muscle cells attached to the surface of a unique type of degradable polymer microsphere are more effective at restoring injured muscle compared with conventional methods of muscle cell delivery. The aim will be achieved through an interdisciplinary project involving biologists, clinicians and materials scientists. The experimental study will include investigating the optimal conditions for attachment of muscle cells to the microspheres; delivery of the cellularized microspheres to pre-clinical models of muscle injury; measurement of integration of the transplanted cells at the injury site; and ability of the transplanted cells to restore muscle contractility. In parallel to the experimental study we will establish women's views on the innovative therapy we are proposing, which will help us to design a therapeutic system best suited to the user's needs.

The outcome from the present study will provide timely, key information for the future development of the therapeutic system. The experimental outputs will be used to design future clinical trials investigating the safety and effectiveness of the treatment in faecally incontinent patients. Both the polymer microspheres and cell types being investigated are already being developed independently for other clinical uses, making transfer of findings from this project to the clinic for faecal incontinence much faster. If successful, the therapeutic system will have tremendous economic benefits to the UK NHS, as well as delivering social, economic and psychological benefits to patients.

Technical Summary

The project aims to demonstrate proof of principle for an innovative therapeutic system for functional restoration of injured muscle using a combination of cell therapy and a novel degradable cell microcarrier device. The novelty of this approach is the use of TIPS microspheres for cell expansion and delivery, utilizing their advantageous features and avoiding the need for proteolytic cell detachment prior to delivery.

Optimal conditions for muscle progenitor cell attachment to TIPS microspheres with different porosities will be identified with primary cultures of myoblasts and mesoangioblasts derived from mouse and human tissues at different seeding densities using a range of dynamic and/or static regimens. External anal sphincter injury will be modelled in a pilot investigation by creating a transverse incision in the tibialis anterior of mice and evaluating the healing response. Delivery of the cellularized microspheres and repair of muscle injury will be investigated in three injury groups simulating repair at different stages after obstetric injury and compared with delivery of cell suspensions or microspheres alone. The muscle will be resected at 2 and 4 weeks after treatment and contractile properties measured using an organ bath and electrical field stimulation. Integration of transplanted cells with host muscle at the injury site and remodelling of tissue will be determined by histology combined with morphometric analysis of fibrosis and angiogenesis.

The findings from the experimental component of the project, combined with feedback from patient user groups, will contribute to the developmental pathway of the therapeutic system. The use of quality qualified TIPS microspheres combined with cell types currently being trialled in humans as advanced therapeutic medicinal products will accelerate future translation of the system into the clinic.

Planned Impact

The project aims to enhance development and translation of a new treatment strategy to the clinic. The project will provide impact in several of the MRC strategic priority areas including delivery of new regenerative medicine products for treatment of disease (Research Priority Theme One; Strategic Aim Two), generation of positive economic impact (Strategic Aim One), and strengthening a skilled research workforce (Strategic Aim Four).

In the short-term (1-3 years), beneficiaries will include companies in the cell therapy industry, especially those that are beginning to explore the untapped market for faecal incontinence (FI). The cell types being investigated are already used as advanced therapeutic medicinal products in commercially-sponsored clinical trials for a variety of clinical conditions. However, existing pre-clinical and clinical data indicate improved methods are sought for delivery of cells. Successful outcomes from this project, combined with TIPS microspheres being quality qualified independently, will deliver a pre-clinically qualified device in readiness for partnership or out-licensing to cell therapy companies for clinical validation of an innovative product in humans and help foster UK economic competitiveness.

In the mid-term (3-5 years), the aim is to verify efficacy of the therapeutic system for delivering cells for the treatment of FI. Successful clinical validation of the therapeutic system will reduce the current economic and social burden of incontinence on the UK NHS and patient community. Although the primary beneficiaries from this project will be patients with FI caused by non-neurogenic damage to anal sphincter muscle, other beneficiaries of the technology will include patients with FI caused by primary degeneration of sphincter muscle. In addition to FI, the therapeutic system will also provide technology of potential value to other conditions associated with muscle insufficiency or where delivery of cells on TIPS microcarriers may prove advantageous e.g. Duchenne muscular dystrophy. Therefore, the intellectual property relating to further development of the device holds significant value for commercialisation. Dissemination of the project findings will help underpin a drive for innovative cell-based therapeutic interventions for this and other clinical conditions, thus boosting the UK economy.

In the longer-term (>5 years), after efficacy for repair of overt sphincter damage with the therapeutic system has been demonstrated, it is feasible to envisage it being applied to the prevention of incontinence in patients who have detectable sphincter disruption (based on future wider adoption of improved ultrasound imaging technology), but who do not currently experience symptoms. The patient perception of this type of approach will be ascertained from the patient participant focus group in the current project. Combined with clinical input for use, these data will influence design of the system and contribute to it meeting requirements of regulatory authorities.

The project will provide the PDRA with an excellent opportunity to develop career skills in a multidisciplinary environment at the laboratory-clinic interface, as well as gaining an insight into the translation pathway from bench-to-bedside. The project will equip the person with the breadth of skills needed to effectively understand the processes involved with developing new healthcare products and provide them with experience of how to establish links and subsequently work with industrial partners as the technology moves forward. These career development opportunities go beyond those typically offered in a pre-clinical research project and will provide the PDRA with key attributes highly sought after for academic-industrial partnerships, adding a skilled scientist to the burgeoning UK med-tech industry work force.

Publications

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Maffioletti SM (2014) Stem cell transplantation for muscular dystrophy: the challenge of immune response. in BioMed research international

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Tedesco FS (2015) Human artificial chromosomes for Duchenne muscular dystrophy and beyond: challenges and hopes. in Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology

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Wright B (2015) A simple and robust method for pre-wetting poly (lactic-co-glycolic) acid microspheres. in Journal of biomaterials applications

 
Title Sculptural works around regenerative medicine technologies 
Description Elpida Hadzi-Vasileva is a contemporary artist who has produced a series of sculptural works around regenerative medicine technologies, including the TIPS microparticles (http://elpihv.co.uk/works/making-beauty2 - slide show images 12 & 13) 
Type Of Art Artwork 
Year Produced 2016 
Impact Elpida Hadzi-Vasileva is a contemporary artist who has produced a series of sculptural works around regenerative medicine technologies, including the TIPS microparticles. 
URL http://elpihv.co.uk/works/making-beauty2
 
Description A Novel Drug-Device Adjuvant Treatment for Radical Prostatectomy
Amount £194,946 (GBP)
Funding ID A22105 
Organisation Cancer Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2017 
End 06/2019
 
Description BHF PhD studentship TIPS microspheres in cardiovascular disease
Amount £70,000 (GBP)
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2014 
End 10/2018
 
Description Biomaterial-Based Therapeutic Neovascularization
Amount £204,681 (GBP)
Funding ID PG/16/56/32246 
Organisation British Heart Foundation (BHF) 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2017 
End 06/2019
 
Description Particle Delivery of Oxidative Biocides for Therapeutic Purposes
Amount £280,000 (GBP)
Organisation Gamma Healthcare 
Sector Private
Country Unknown
Start 10/2016 
End 01/2017
 
Description Proof-of-concept award
Amount £25,000 (GBP)
Organisation UCL Business 
Sector Private
Country United Kingdom
Start 11/2014 
End 03/2015
 
Description Refined Manufacturing and Process Development of TIPS Particle Technology
Amount £110,000 (GBP)
Organisation National Institute for Health Research 
Department UCLH/UCL Biomedical Research Centre
Sector Public
Country United Kingdom
Start 12/2014 
End 12/2016
 
Description Refined Manufacturing and Process Development of TIPS Particle Technology
Amount £183,236 (GBP)
Organisation National Institute for Health Research 
Department UCLH/UCL Biomedical Research Centre
Sector Public
Country United Kingdom
Start 03/2016 
End 11/2017
 
Description Regenerative Medicine Research Committee
Amount £214,234 (GBP)
Funding ID MR/R014108/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 12/2018 
End 01/2019
 
Description TIPS microspheres for a novel antimicrobial release system aimed at tackling antimicrobial resistance
Amount £60,000 (GBP)
Organisation Medical Research Council (MRC) 
Department MRC Confidence in Concept Scheme
Sector Academic/University
Country United Kingdom
Start 09/2014 
End 09/2015
 
Description TIPS microspheres in cardiovascular disease
Amount £20,000 (GBP)
Organisation Bristol Heart Institute 
Sector Charity/Non Profit
Country United Kingdom
Start 07/2014 
End 07/2015
 
Description UK Regenerative Medicine Platform Hub Partnership Award
Amount £65,642 (GBP)
Organisation UKRMP Acellular Hub and UKRMP Immunomodulation Hub (Delcassian) 
Sector Academic/University
Country United Kingdom
Start 10/2014 
End 08/2017
 
Title Patient and public perceptions of novel regenerative medicine therapies. 
Description We have devised a novel questionnaire to gain perceptions on the proposed novel therapy involving cell therapy and regenerative medicine. This involves patient and public involvement (PPI) and includes collaboration between clinical teams at UCL H. 
Type Of Material Improvements to research infrastructure 
Year Produced 2014 
Provided To Others? Yes  
Impact We have completed collecting the data with the questionaaire for analysis. The results have provided a clear insight into how the general public perceive and understand our research. The results have now been published: • Wright B, Emmanuel A, Athanasakos E, Parmar N, Parker G, Green B, Tailby E, Chandler H, Cushnie J, Pembroke J, Saruchera Y, Vashisht A, Day R. (2016) Women's Views on Autologous Cell-Based Therapy for Post-Obstetric Incontinence. Regenerative Medicine (in press). doi:10.2217/rme.15.88 
 
Title Precision attachment & storage of cells on microcarriers 
Description Recent advances in biotechnology have introduced the possibility of new therapies based on either the delivery of cells or isolation of their biological derivatives. Adherent cells are those that exist in a native state attached to surrounding tissues. When isolated from their normal environment adherent cells require an artificial substrate in order to grow and divide in vitro. Adherent cells are being explored as novel therapies for a variety of conditions as well for drug discovery as replacements for animal models to screen new compounds, or for the production of biological materials, such as enzymes, vaccines, hormones, antibodies, interferons and nucleic acids. Conventional methods used for manufacturing and storage of adherent cells are sub-optimal for many of the new uses, where delivery of precise quantities of viable and potent cells is often required. Microcarriers are being increasingly explored for the expansion of adherent cells as an alternative to conventional 2D tissue culture substrates. However, two interlinked stages frequently used during the manufacturing process of adherent cell products pose particular risks to the viability and potency of the product: The first risk occurs if there is a need for initial detachment of cells from the culture substrate, which poses a critical risk to the viability of the cells and their ultimate function. The second risk occurs if the cells require cryopreservation during the transfer from the manufacturer to the end user, with cryopreservation and cell thawing having an impact on cell survival. To address these challenges we have developed novel clinically ready, biodegradable, biocompatible synthetic microcarriers (TIPS microparticles) that eliminate the need for cell detachment before clinical delivery. Our pre-clinical studies have demonstrated this approach is technically feasible and the technology is currently being translated to early stage clinical studies. Building on this, our pilot studies have demonstrated that cells attached to the biodegradable microcarriers are compatible with cryopreservation, with viable cells remaining attached to the microcarriers after thawing. We hypothesize that the microcarriers offer a feasible solution to mitigate the risks associated with the manufacture of adherent cell based products used in biotechnology. Since existing methods for seeding adherent cells onto microcarriers are unpredictable and inconsistent we have devised a novel process that involves incubating individual microcarriers with a specific quantity of cells in a droplet of liquid. To date, our pilot studies have demonstrated this approach is technically feasible but have involved the use of manual dispensing of cells and microcarriers into the hanging drop. To make the technology attractive for industrial applications, we hypothesize this approach could be scaled-up and automated to provide a process capable of producing large quantities of cellularized microcarriers, each with a specified quantity of cells attached to the surface. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact We are in the process of filing a patent application on the technology outlined above. 
 
Description Force Generation Studies with Prof Dominic Wells at Royal Veterinary College 
Organisation Royal Veterinary College (RVC)
Country United Kingdom 
Sector Academic/University 
PI Contribution We have developed an in vivo injury model designed to assess the potency of the advanced therapeutic medicinal products being developed.
Collaborator Contribution A method for evaluating the regeneration of muscle, in terms for force generation, has been validated for use with the advanced therapeutic medicinal products being developed.
Impact The collaboration is multi-disciplinary, involving cell biology, materials science and surgery.
Start Year 2014
 
Description Investigation of muscle regenerative therapies 
Organisation Cook Myosite Inc
Country United States 
Sector Private 
PI Contribution Evaluation of compatibility of Cook Myosite cell products and reagents with our cell manufacturing technology.
Collaborator Contribution Provision of research materials and insight into industry requirements.
Impact Multidisciplinary collaboration between cell biologists, biomedical engineers and bioprocessing experts.
Start Year 2016
 
Description Investigation of muscle regenerative therapies 
Organisation Merck
Department MilliporeSigma
Country United States 
Sector Private 
PI Contribution Development of novel manufacturing processes for cell based medicines.
Collaborator Contribution Insight into industry requirements.
Impact Multidisciplinary collaboration between cell biologists, biomedical engineers and bioprocessing experts.
Start Year 2016
 
Description Investigation of muscle regenerative therapies 
Organisation Plasticell Ltd
Country United Kingdom 
Sector Private 
PI Contribution Provision of research materials to evaluate their potential use in controlled delivery of active ingredients.
Collaborator Contribution Verification of human induced pluripotent stem cell attachment to TIPS microparticles.
Impact A collaborative project with Plasticell Ltd, an SME based at the Stevenage Bioscience Catalyst site, is investigating the use of TIPS microspheres to assist with their proprietary technology differentiating human induced pluripotent stem cells into myogenic progenitors and the controlled delivery of inducers of muscle differentiation they have discovered through screening of GSK small molecule libraries. A joint funding application has entered the second stage of the Technology Strategy Board's Advancing Regenerative Medicines and Cell Therapies scheme.
Start Year 2014
 
Description Research Collaboration with Dr Daniel Stuckey at UCL Centre for Advanced Biomedical Imaging 
Organisation University College London
Department Department of Geography
Country United Kingdom 
Sector Academic/University 
PI Contribution Access to research models that highlight the value of advanced biomedical imaging modalities.
Collaborator Contribution The new collaboration has enabled the application of non-destructive imaging and tracking of cellularized biomaterial constructs being explored in our research.
Impact Research publication being prepared.
Start Year 2015
 
Description All Party Parliamentary Group for Continence Care,House of Lord's (London, UK) 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Discussion on continence care and how new technologies including regenerative medicine might benefit this.
Year(s) Of Engagement Activity 2014
 
Description EDANA (international association for the nonwovens and related industries) Outlook conference (Barcelona, Spain) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Invited lecture on biomaterials and incontinence. Established new links with industry.
Year(s) Of Engagement Activity 2014
 
Description Invited Talk Tissue and Cell Engineering Society/UK Regenerative Medicine Platform Hub Meeting, Newcastle, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Invited talk on TIPS microsphere technology. Further collaborations established using TIPS microspheres.
Year(s) Of Engagement Activity 2014
 
Description Key note lecture TERM STEM 2015 (Guimarães, Portugal) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact To present data from project and engage with fellow researchers. New collaborations have arisen from this activity.
Year(s) Of Engagement Activity 2015
 
Description Speaker at Inaugural UK Regenerative Medicine Conference 20 / 21 September 2016 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Follow-up discussions with academics and industry that have led to new collaborations.
Year(s) Of Engagement Activity 2016
 
Description UCB Discovery Biology Science Day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Keynote lecture on the TIPS microparticle technology at the UCB Discovery Biology Science Day. UCB is a global biopharmaceutical company focusing on creating value for people living with neurology and immunology conditions.
Year(s) Of Engagement Activity 2016