Bioengineering Thymus Organoids to Restore Thymic Function and Induce Donor-Specific Immune Tolerance to Allografts

Fan, Yong; Tajima, Asako; Goh, Saik Kia; Geng, Xuehui; Gualtierotti, Giulio; Grupillo, Maria; Coppola, Antonina; Bertera, Suzanne; Rudert, William A.; Banerjee, Ipsita; Bottino, Rita; Trucco, Massimo

doi:10.1038/mt.2015.77

Cited by 99 publications

(101 citation statements)

References 70 publications

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“…In this technique, all cells are removed from the organ, leaving the extracellular matrix intact. It has been reported that decellularization of the thymus, followed by reconstitution with thymic stromal cells and lineage negative BM progenitors, led to formation a functional thymus when transplanted into the kidney capsule of nude mice (192). …”

Section: Exogenous Strategies To Enhance Thymic Recoverymentioning

confidence: 99%

Thymus: the next (re)generation

Chaudhry

Velardi

Dudakov

et al. 2016

Immunological Reviews

145

131

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Summary As the primary site of T cell development, the thymus plays a key role in the generation of a strong yet self-tolerant adaptive immune response, essential in the face of the potential threat from pathogens or neoplasia. As the importance of the role of the thymus has grown, so too has the understanding that it is extremely sensitive to both acute and chronic injury. The thymus undergoes rapid degeneration following a range of toxic insults, and also involutes as part of the aging process, albeit at a faster rate than many other tissues. The thymus is, however, capable of regenerating, restoring its function to a degree. Potential mechanisms for this endogenous thymic regeneration include keratinocyte growth factor (KGF) signaling, and a more recently described pathway in which innate lymphoid cells produce interleukin-22 (IL-22) in response to loss of double positive thymocytes and upregulation of IL-23 by dendritic cells. Endogenous repair is unable to fully restore the thymus, particularly in the aged population, and this paves the way towards the need for exogenous strategies to help regenerate or even replace thymic function. Therapies currently in clinical trials include KGF, use of the cytokines IL-7 and IL-22, and hormonal modulation including growth hormone administration and sex steroid inhibition. Further novel strategies are emerging in the pre-clinical setting, including the use of precursor T cells and thymus bioengineering. The use of such strategies offers hope that for many patients, the next regeneration of their thymus is a step closer.

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Section: Exogenous Strategies To Enhance Thymic Recoverymentioning

confidence: 99%

Thymus: the next (re)generation

Chaudhry

Velardi

Dudakov

et al. 2016

Immunological Reviews

145

131

View full text Add to dashboard Cite

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“…Monitor the development of T cells in the periphery by harvesting 50–100 µl blood samples into the EDTA-containing blood collection tube from the nude mice 4–6 weeks post transplantation and stain with a cocktail of anti-CD4, -CD8, -CD45, and -CD3 antibodies 12 .…”

Section: Protocolmentioning

confidence: 99%

“…Analyze the percentages of T-lymphocytes in the peripheral blood leukocytes with flow cytometry (FCM), using single cell analysis software 12 .…”

Section: Protocolmentioning

confidence: 99%

“…Using similar technique, we have recently demonstrated that 3-D aggregation of TECs could be promoted in EAK hydrogel with adaptor complexes carrying TEC-specific anti-EpCAM antibodies (αH6:pA/G:αEpCAM). When transplanted underneath the kidney capsules of nude mice, the TEC clusters embedded in EAK hydrogel can effectively support the development of functional T cells in vivo 11,12 .…”

Section: Introductionmentioning

confidence: 99%

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Promoting 3-D Aggregation of FACS Purified Thymic Epithelial Cells with EAK 16-II/EAKIIH6 Self-assembling Hydrogel

Tajima

Liu

Pradhan

et al. 2016

JoVE

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Thymus involution, associated with aging or pathological insults, results in diminished output of mature T-cells. Restoring the function of a failing thymus is crucial to maintain effective T cell-mediated acquired immune response against invading pathogens. However, thymus regeneration and revitalization proved to be challenging, largely due to the difficulties of reproducing the unique 3D microenvironment of the thymic stroma that is critical for the survival and function of thymic epithelial cells (TECs). We developed a novel hydrogel system to promote the formation of TEC aggregates, based on the self-assembling property of the amphiphilic EAK16-II oligopeptides and its histidinylated analogue EAKIIH6. TECs were enriched from isolated thymic cells with density-gradient, sorted with fluorescence-activated cell sorting (FACS), and labeled with anti-epithelial cell adhesion molecule (EpCAM) antibodies that were anchored, together with anti-His IgGs, on the protein A/G adaptor complexes. Formation of cell aggregates was promoted by incubating TECs with EAKIIH6 and EAK16-II oligopeptides, and then by increasing the ionic concentration of the medium to initiate gelation. TEC aggregates embedded in EAK hydrogel can effectively promote the development of functional T cells in vivo when transplanted into the athymic nude mice.

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“…Limited, but encouraging functional recoveries of the engineered organoids have been observed in preclinical studies[19, 20]. Taking advantage of the tissue engineering approach, we have reconstructed thymus organoids by repopulating decellularized thymus scaffolds with isolated TECs[21]. The thymus organoids can support T-cell development both in vitro and in vivo .…”

Section: Introductionmentioning

confidence: 99%

Construction of Thymus Organoids from Decellularized Thymus Scaffolds

Tajima

Pradhan

Geng

et al. 2016

Methods in Molecular Biology

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Summary One of the hallmarks of modern medicine is the development of therapeutics that can modulate immune responses, especially the adaptive arm of immunity, for disease intervention and prevention. While tremendous progresses have been made in the past decades, manipulating the thymus, the primary lymphoid organ responsible for the development and education of T lymphocytes, remains a challenge. One of the major obstacles is the difficulty to reproduce its unique extracellular matrix (ECM) microenvironment that is essential for maintaining the function and survival of thymic epithelial cells (TECs), the predominant population of cells in the thymic stroma. Here, we described the construction of functional thymus organoids from decellularized thymus scaffolds repopulated with isolated TECs. Thymus decellularization was achieved by freeze/thaw cycles to induce intracellular ice crystal formation, followed by detergent-induced cell lysis. Cellular debris was removed with extensive wash. The decellularized thymus scaffolds can largely retain the 3-D extracellular matrix (ECM) microenvironment that can support the re-colonization of TECs. When transplanted into athymic nude mice, the reconstructed thymus organoids can effectively promote the homing of bone marrow-derived lymphocyte progenitors and support the development of a complex T cell repertoire. Bioengineering of thymus organoids can be a promising approach to rejuvenate/modulate the function of T-cell mediated adaptive immunity in regenerative medicine.

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Bioengineering Thymus Organoids to Restore Thymic Function and Induce Donor-Specific Immune Tolerance to Allografts

Cited by 99 publications

References 70 publications

Thymus: the next (re)generation

Thymus: the next (re)generation

Promoting 3-D Aggregation of FACS Purified Thymic Epithelial Cells with EAK 16-II/EAKIIH6 Self-assembling Hydrogel

Construction of Thymus Organoids from Decellularized Thymus Scaffolds

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