The Impact of Cell and Gene Therapy on the Function and Molecular Regulation of Hematopoietic Stem Cells.
Lead Research Organisation:
University of Cambridge
Department Name: Wellcome Trust - MRC Cam Stem Cell Inst
Abstract
Haematopoiesis is an intricate cellular process allowing the generation and further maintenance of a diverse repertoire of functional blood cells
throughout an organisms' lifetime. Hematopoietic stem cells (HSCs) constitute the foundation of the hematopoietic hierarchy and have the potential to
both self-renew and produce lineage specific progenitors. The fundamental properties of HSCs to expand and differentiate have provoked interest in
their application in gene therapy for the treatment of a range of hematopoietic diseases. However, the molecular consequences of gene therapy on the
underlying biology of HSCs remains poorly understood. This project aims to address unanswered areas in the field by combining single cell
transcriptomics, lentiviral transduction technology, flow cytometry and single cell functional assays. Importantly, research conducted will both improve
our overall understanding of how HSCs respond to stress and help to determine whether gene therapy is a safe and effective means of treatment.
Results obtained from this project also have the potential to guide future improvements to the gene therapy protocol in order to increase HSC
functionality after transduction.
throughout an organisms' lifetime. Hematopoietic stem cells (HSCs) constitute the foundation of the hematopoietic hierarchy and have the potential to
both self-renew and produce lineage specific progenitors. The fundamental properties of HSCs to expand and differentiate have provoked interest in
their application in gene therapy for the treatment of a range of hematopoietic diseases. However, the molecular consequences of gene therapy on the
underlying biology of HSCs remains poorly understood. This project aims to address unanswered areas in the field by combining single cell
transcriptomics, lentiviral transduction technology, flow cytometry and single cell functional assays. Importantly, research conducted will both improve
our overall understanding of how HSCs respond to stress and help to determine whether gene therapy is a safe and effective means of treatment.
Results obtained from this project also have the potential to guide future improvements to the gene therapy protocol in order to increase HSC
functionality after transduction.
| Description | Improving gene therapy protocol efficacy by modulating haematopoietic stem cell exit from quiescence |
| Amount | £71,077 (GBP) |
| Organisation | University of Cambridge |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 01/2018 |
| End | 10/2018 |
| Description | Industrial contribution to "The Impact of Cell and Gene Therapy on the Function and Molecular Regulation of Hematopoietic Stem Cells." |
| Amount | £119,914 (GBP) |
| Organisation | GlaxoSmithKline (GSK) |
| Sector | Private |
| Country | Global |
| Start | 10/2017 |
| End | 01/2022 |
| Description | Leveraging mRNA technology to improve haematopoietic stem cell functional fitness during the ex vivo gene therapy protocol |
| Amount | £275,272 (GBP) |
| Organisation | CSL Behring, Switzerland |
| Sector | Private |
| Country | Switzerland |
| Start | 06/2023 |
| End | 05/2025 |
| Title | Characterization of transcriptomic changes induced by a gene therapy cell culture protocol to single human bone marrow and mobilized peripheral blood hematopoietic stem and progenitor cells |
| Description | Here we present the single cell transcriptomes of human adult mobilized peripheral blood (mPB) and bone marrow (BM) hematopoietic stem and progenitor cell (HSPC) subsets,: long-term haematopoietic stem cells (LT-HSCs), short-term HSCs (ST-HSCs) and CD34+ cells before (0 h) and after 62 hours (62 h) of culture in a conditions mimicking gene therapy protocols. The culture protocol includes two hits of lentiviral transduction (lentiviral vector contains a GFP construct) and media as previously published . |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| Impact | No impact yet |
| URL | https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE213370 |
| Title | Time-course analysis of transcriptome changes induced by ex vivo culture in human cord blood hematopoietic stem cell at single cell resolution |
| Description | Here we present the single cell transcriptomes of human cord blood (CB) long-term hematopoietic stem cells (LT-HSCs) cultured in differentiation promoting medium over a time-course of 0, 6 24 or 72 h. A condition was included for LT-HSCs cultured with the CDK6 inhibitor Palbociclib for durations of 24 or 72 h. Palbociclib causes reversible cell cycle arrest in early G1 in these culture conditions. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| Impact | No impact yet |
| URL | https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE213365 |
| Description | Collaboration with Cell & Gene Therapy Department at GSK |
| Organisation | GlaxoSmithKline (GSK) |
| Department | GlaxoSmithKline, Stevenage |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | I generated a CITE-Seq time-course dataset created to investigate the lag phase of T cell expansion. |
| Collaborator Contribution | Assistance with generation of CITE-Seq libraries for sequencing and analysis files. |
| Impact | No outcomes yet. |
| Start Year | 2021 |
| Description | Collaboration with Dr Cabezas-Wallscheid, Max Planck Institute, Freiburg, Germany |
| Organisation | Max Planck Society |
| Department | Max Planck Institute of Immunobiology and Epigenetics |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | During this collaboration, we have used our expertise in human haematopoietic stem cell quiescence to a study that describes the first marker for human dormant haematopoietic stem cells. Our data has been incorporated into a study currently (Dec 2021) under revision at Nature Cell Biology. |
| Collaborator Contribution | We exchanged information on human haematopoietic stem cell quiescence. |
| Impact | One publication where I am co-author published in Nature Cell Biology (10.1038/s41556-022-00931-x) |
| Start Year | 2019 |
| Description | Collaboration with Dr Kent laboratory on ex vivo hibernation of hematopoietic stem cells |
| Organisation | University of York |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Based on the expertise developped in this award we performed experiments aimed at establishing a culture system to maintain human haematopoietic stem cells in a viable, reversible non dividing state ex vivo. |
| Collaborator Contribution | The Kent lab had established similar culture conditions for mouse haematopoietic stem cells. The information they had gathered on how cells behave in this culture was very useful and complementary to the data generated in this award. |
| Impact | In October 2020, we submitted a joint paper for publication which was eventually accepted at Stem Cell Reports and published in June 2021. |
| Start Year | 2017 |
| Description | Collaboration with Prof. Green research group on understanding MHC Class II expression in cultured human haematopoietic stem cells |
| Organisation | University of Cambridge |
| Department | Cambridge Stem Cell Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | During this collaboration, we re-analysed a published single cell RNA-Seq dataset to show that expression of a subset of MHC Class II molecules is higher in a subset of Haematopoeitic Stem Cells (HSCs) defined by slow cell cycle kinetics and multipotent lineage output. This data in the human system was incorperated into a manuscript written by the Green group which is in revision at Blood. |
| Collaborator Contribution | Analysis of published single cell RNA-Seq dataset |
| Impact | Manuscript in revision at Blood. |
| Start Year | 2021 |
| Description | Human HSC Dormancy Marker Investigation |
| Organisation | Max Planck Society |
| Department | Max Planck Institute of Immunobiology and Epigenetics |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | The collaborating laboratory in Germany is investigating a novel cell surface marker to enrich for a population of dormant human haematopoietic stem cells (HSCs). Our laboratory in Cambridge has established protocols for investigation of in vitro single cell differentiation capacity in a high throughput manner. We collaborated on this project by performing and analysing results from single cell differentiation assays to determine the proliferation and differentiation output of single human HSCs expressing this novel marker. |
| Collaborator Contribution | The collaborating laboratory provided the details of the novel marker and guidance with analysis. |
| Impact | The results from the experiments performed have been included in a manuscript that has been submitted. |
| Start Year | 2020 |
| Description | Blood Stem Cell Robots |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | With the BBSRC funding associated with this award, we have designed an interactive activity aimed at introducing children to how different blood cell types are formed from blood stem cells. To do this, we used a set of small Thymio robot 'stem cells' which children programme to navigate a series of decision branches on a large floor mat to reach their chosen cell type. Researchers have first explained how they study this stem cell behavior in the laboratory and then have helped the children to run the activity. This event was run in the following settings: i) 2017: Cherry Hinton library; attracted approximately 30 children and their parents. ii) 2018: a) School visit to Primary School in Saffron Waldon organised with PluriMes. Full day of rotational class activities, one of which was Stem Cell Robots. b) School visits to two under-served Cambridge Primary Schools as part of UniStemDay. 1 hour class group sessions with Stem Cell Robots. c) Action Stations - Centenary celebration day at Homerton College for college alumni and members of the public. d) Family STEAM day at the Spinney School for local families. e) Life Lab - European Researchers Night - Science in the Square. Weekday festival tent in Peterborough Shopping Centre for under-served local community. iii) 2019: Life Lab - European Researchers Night. Weekend festival tent at Ely Cathedral for local families. iv) 2020: a) Girls in Stem Day - Ely College. Rotating hands on activity sessions for girl guiding groups from across the county. |
| Year(s) Of Engagement Activity | 2017,2018,2019,2020 |
| Description | Interview for Italian website 30Science.com |
| Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | I provided an interview for the Italian website 30Science.com following receipt of my ERC Consolidator grant. |
| Year(s) Of Engagement Activity | 2023 |
| URL | https://30science.com/2023/02/in-evidenza/laurenti/ |
| Description | Primary School Visit (Cambridge) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Visited a local primary school to hold a workshop for 8-9 year old pupils surrounding the theme of blood stem cells. Here, I explained the concept of stem cell dormancy and cell fate decisions in an age appropriate manner and a short presentation was given on the types of mature cells found in human blood. The students then participated in an interactive board game where they could simulate the "decisions" a hematopoietic stem cell makes to eventually become a mature blood cell. Here, I helped the students with the activity and answered any questions they had about blood/ cell biology. I then showed the students microscopy images of mature blood cells and assisted the students with crosswords containing key words relevant to stem cell biology. Throughout the workshop I also answered any questions they had regarding working in a laboratory and being a scientist. The feedback that we received from the school stated that the students enjoyed the workshop and suggested that the Cambridge Stem Cell Institute return for future school visits. I thoroughly enjoyed leading the session and hope to participate again in future school visits. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Public Engagement at Cambridge College |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Participated in a science festival showcasing the Cambridge Stem Cell Institute as part of the Homerton College (Cambridge) 250th anniversary celebrations. Here, the public took part in an activity whereby volunteers could simulate the decisions that a stem cell makes in order to become a mature cell. I was then able to speak to individuals and give an insight into my PhD project investigating gene therapy in hematopoietic stem cells. The public seemed engaged in the activity and had many questions surrounding my PhD project. It was interesting to hear the publics prior perceptions on gene therapy and many were aware of the potential use of CRISPR technologies in future healthcare. I also enjoyed talking to the public about the current application of gene therapy in the context of treating inherited blood disorders such as primary immunodeficiencies, and the difficulties being faced with this approach. Many of the public were unaware of the recent scientific advances that have led to the approval of this therapy in Europe and seemed interested in the discussion. |
| Year(s) Of Engagement Activity | 2018 |
| URL | https://homerton250.org/events/festival/ |
| Description | Selected Speaker at Homerton College Research Supper |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | Selected speaker at the Homerton Research supper delivering a 20 minute talk to members of the college across all disciplines. Following the talk I received many questions about gene therapy and it's potential to treat genetic disease. Following my talk there was a discussion session which was well attended and led to an interesting conversation on the future of healthcare. |
| Year(s) Of Engagement Activity | 2021 |