Difference between revisions of "Part:BBa K2549001"

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CD19-targeted chimeric antigen receptor T-cell therapy<ref>CD19-targeted chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Maude SL, Teachey DT, Porter DL, Grupp SA. Blood, 2015 Jun;125(26):4017-23  PMID: 25999455; DOI: 10.1182/blood-2014-12-580068</ref>: Acute lymphoblastic leukemia (ALL) remains difficult to treat, with minimal improvement in more than 2 decades. Adoptive transfer of T cells engineered to express a chimeric antigen receptor (CAR) has emerged as a powerful targeted immunotherapy. Complete remission rates as high as 90% have been reported in children and adults with relapsed and refractory ALL treated with CAR-modified T cells targeting the B-cell–specific antigen CD19. For more details, please check '''Maude SL''' et al.
 
CD19-targeted chimeric antigen receptor T-cell therapy<ref>CD19-targeted chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Maude SL, Teachey DT, Porter DL, Grupp SA. Blood, 2015 Jun;125(26):4017-23  PMID: 25999455; DOI: 10.1182/blood-2014-12-580068</ref>: Acute lymphoblastic leukemia (ALL) remains difficult to treat, with minimal improvement in more than 2 decades. Adoptive transfer of T cells engineered to express a chimeric antigen receptor (CAR) has emerged as a powerful targeted immunotherapy. Complete remission rates as high as 90% have been reported in children and adults with relapsed and refractory ALL treated with CAR-modified T cells targeting the B-cell–specific antigen CD19. For more details, please check '''Maude SL''' et al.
  
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===== Reported data from Royal KT et al 2016=====
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===== Works extremely well in Royal KT et al 2016=====
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Please refer the original article for more details<ref>Engineering T Cells with Customized Therapeutic Response Programs Using Synthetic Notch Receptors. Roybal KT, Williams JZ, Morsut L, ..., McNally KA, Lim WA. Cell, 2016 Oct;167(2):419-432.e16  PMID: 27693353; DOI: 10.1016/j.cell.2016.09.011</ref>. Below is our summary of their article to explain why we focus on CD19.
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Please refer the original article for more details<ref>Engineering T Cells with Customized Therapeutic Response Programs Using Synthetic Notch Receptors. Roybal KT, Williams JZ, Morsut L, ..., McNally KA, Lim WA. Cell, 2016 Oct;167(2):419-432.e16  PMID: 27693353; DOI: 10.1016/j.cell.2016.09.011</ref>. Below is our summary of their article to explain '''why we focus on CD19'''.
  
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[[File:CD19a.png|1800px|CD19 related data in Royal KT 2016]]
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[[File:CD19c.png|360px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated: ''SynNotch receptors have a custom ligand binding domain that detects a cell-surface antigen of interest (e.g., scFvs targeted to CD19 or Her2 or nanobodies to GFP), the core regulatory region of Notch that controls proteolysis, and a cytoplasmic orthogonal transcription factor (e.g., Gal4 VP64). The corresponding response elements for the orthogonal transcription factor controlling custom transcriptional programs are also engineered into the T cell.''
  
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[[File:CD19b.png|360px|CD19 related data in Royal KT 2016]]
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[[File:CD19a.png|180px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated: ''CD4+ and CD8+ primary human T cells were engineered with the a-CD19 synNotch Gal4VP64 receptor and 5x Gal4 response elements control- ling the expression of a BFP reporter. Histogram showing selective induction of the BFP reporter in a-CD19 synNotch receptor receiver CD4+ T cells in response to stimulation with sender cells with CD19� or CD19+ K562s.''
  
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[[File:CD19c.png|360px|CD19 related data in Royal KT 2016]]
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[[File:CD19b.png|360px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated: ''CD4+ AND CD8+ primary human T cells were engineered with the a-CD19 nanobody synNotch Gal4VP64 receptors and 5x Gal4 response elements controlling the expression of a BFP reporter. The percentages of synNotch T cells that upregulate the BFP reporter after 24 hr of stimulation with the indicated sender cells is given (n >= 3 for all conditions, error bars, SEM).''
  
 
[[File:CD19d.png|180px|CD19 related data in Royal KT 2016]]
 
[[File:CD19d.png|180px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
 
[[File:CD19e.png|360px|CD19 related data in Royal KT 2016]]
 
[[File:CD19e.png|360px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
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[[File:CD19f.png|360px|CD19 related data in Royal KT 2016]]
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[[File:CD19f.png|540px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
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[[File:CD19g.png|360px|CD19 related data in Royal KT 2016]]
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[[File:CD19g.png|180px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
 
[[File:CD19h.png|360px|CD19 related data in Royal KT 2016]]
 
[[File:CD19h.png|360px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
 
[[File:CD19i.png|360px|CD19 related data in Royal KT 2016]]
 
[[File:CD19i.png|360px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
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[[File:CD19j.png|360px|CD19 related data in Royal KT 2016]]
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[[File:CD19j.png|180px|CD19 related data in Royal KT 2016]]
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Royal KT at al stated:
  
 
===== Works as we designed =====
 
===== Works as we designed =====

Revision as of 06:29, 7 October 2018


suface-expressed CD19

Surface-expressed CD19 (surCD19) is built by joining CD8α signal peptide, CD19 extracellular domain and the transmembrane region of PDGFRβ (from N terminal to C terminal). Additional HA tag on its N terminal and a Myc tag on its C terminal to facilitate detection by antibodies[1]. CD8α peptide guides synthesized fusion protein to pass the translocon[2] into the endoplasmic reticulum[3], and the fusion protein will be later sugar modified in Golgi[4], presented on the plasma membrane and located to the outside of the cell. Transmembrane region of PDGFRβ embeds surCD19 on the membrane. It was used as the antigen for Part:BBa_K2549005.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 588
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 757
    Illegal SapI site found at 273
    Illegal SapI.rc site found at 91

Biology

Clinical significance of CD19

As summarized on wikipedia page[5]: B-lymphocyte antigen CD19, also known as CD19 molecule (Cluster of Differentiation 19), B-Lymphocyte Surface Antigen B4, T-Cell Surface Antigen Leu-12 and CVID3 is a transmembrane protein that in humans is encoded by the gene CD19. In humans, CD19 is expressed in all B lineage cells, except for plasma cells, and in follicular dendritic cells. CD19 plays two major roles for B cells: (1) it acts as an adaptor protein to recruit cytoplasmic signaling proteins to the membrane; (2) it works within the CD19/CD21 complex to decrease the threshold for B cell receptor signaling pathways. Due to its presence on all B cells, it is a biomarker for B lymphocyte development, lymphoma diagnosis and can be utilized as a target for leukemia immunotherapies.

CD19-targeted chimeric antigen receptor T-cell therapy[6]: Acute lymphoblastic leukemia (ALL) remains difficult to treat, with minimal improvement in more than 2 decades. Adoptive transfer of T cells engineered to express a chimeric antigen receptor (CAR) has emerged as a powerful targeted immunotherapy. Complete remission rates as high as 90% have been reported in children and adults with relapsed and refractory ALL treated with CAR-modified T cells targeting the B-cell–specific antigen CD19. For more details, please check Maude SL et al.

Works extremely well in Royal KT et al 2016

Please refer the original article for more details[7]. Below is our summary of their article to explain why we focus on CD19.

CD19 related data in Royal KT 2016 Royal KT at al stated: SynNotch receptors have a custom ligand binding domain that detects a cell-surface antigen of interest (e.g., scFvs targeted to CD19 or Her2 or nanobodies to GFP), the core regulatory region of Notch that controls proteolysis, and a cytoplasmic orthogonal transcription factor (e.g., Gal4 VP64). The corresponding response elements for the orthogonal transcription factor controlling custom transcriptional programs are also engineered into the T cell.

CD19 related data in Royal KT 2016 Royal KT at al stated: CD4+ and CD8+ primary human T cells were engineered with the a-CD19 synNotch Gal4VP64 receptor and 5x Gal4 response elements control- ling the expression of a BFP reporter. Histogram showing selective induction of the BFP reporter in a-CD19 synNotch receptor receiver CD4+ T cells in response to stimulation with sender cells with CD19� or CD19+ K562s.

CD19 related data in Royal KT 2016 Royal KT at al stated: CD4+ AND CD8+ primary human T cells were engineered with the a-CD19 nanobody synNotch Gal4VP64 receptors and 5x Gal4 response elements controlling the expression of a BFP reporter. The percentages of synNotch T cells that upregulate the BFP reporter after 24 hr of stimulation with the indicated sender cells is given (n >= 3 for all conditions, error bars, SEM).

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

CD19 related data in Royal KT 2016 Royal KT at al stated:

Works as we designed

surCD19

on the membrane, flow and immunostaining


References

  1. ↑ Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Morsut L, Roybal KT, Xiong X, ..., Thomson M, Lim WA. Cell, 2016 Feb;164(4):780-91 PMID: 26830878; DOI: 10.1016/j.cell.2016.01.012
  2. ↑ https://en.wikipedia.org/wiki/Translocon
  3. ↑ https://en.wikipedia.org/wiki/Endoplasmic_reticulum
  4. ↑ https://en.wikipedia.org/wiki/Golgi_apparatus
  5. ↑ https://en.wikipedia.org/wiki/CD19
  6. ↑ CD19-targeted chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Maude SL, Teachey DT, Porter DL, Grupp SA. Blood, 2015 Jun;125(26):4017-23 PMID: 25999455; DOI: 10.1182/blood-2014-12-580068
  7. ↑ Engineering T Cells with Customized Therapeutic Response Programs Using Synthetic Notch Receptors. Roybal KT, Williams JZ, Morsut L, ..., McNally KA, Lim WA. Cell, 2016 Oct;167(2):419-432.e16 PMID: 27693353; DOI: 10.1016/j.cell.2016.09.011