Antigen-mediated migration of murine pro-B Ba/F3 cells via an antibody/receptor chimera

Hitomi

Nagamune

2014

Biotechnology Progress

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Since cell migration plays critical roles in development and homeostasis of the body, artificial control of cell migration would be promising for the treatment of various diseases related to migration. To this end, we previously developed single-chain Fv (scFv)/receptor chimeras, named signalobodies, which can control cell fates via a specific antigen that is different from natural cytokines. Although a conventional chemotaxis chamber assay revealed that several signalobodies based on receptor tyrosine kinases transduced antigen-dependent migration signals, we have never performed direct observation of the cells to obtain more information on overall properties of cell motility and migration. In this study, we utilized murine pro-B Ba/F3 cells expressing either a scFv-Fms or scFv-Mpl signalobody, and compared their migratory characteristics. We employed a lipid-polyethylene glycol conjugate to softly immobilize the suspension cells on a slide, which facilitated direct observation of chemokinetic activity of the cells. Consequently, both cells markedly exhibited chemokinesis in response to a specific antigen. In addition, the cells were subjected to a stable antigen-concentration gradient to observe horizontal directional cell migration in real time. The results showed that the cells expressing scFv-Fms underwent directional migration toward a positive antigen-concentration gradient. Taken together, we successfully demonstrated antigen-responsive regulation of cell motility and migration via the signalobodies.

Section: Resultsmentioning

confidence: 83%

“…Ba/S‐Fms was also subjected to a migration assay using transwell plates. The result indicated that S‐Fms transduces both chemotactic and chemokinetic signals in response to BSA‐FL …”

Section: Resultsmentioning

confidence: 99%

Section: Comparison Of Expression Levels Of S-fms and S-mpl On Ba/f3 mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Antigen‐responsive regulation of Cell motility and migration via the signalobodies based on c‐Fms and c‐Mpl

Hitomi

Nagamune

2014

Biotechnology Progress

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S‐Fms signalobody enhances myeloid cell growth and migration

Hitomi

Nagamune

2014

Biotechnology Journal

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Since receptor tyrosine kinases (RTKs) control various cell fates in many types of cells, mimicry of RTK functions is promising for artificial control of cell fates. We have previously developed single-chain Fv (scFv)/receptor chimeras named signalobodies that can mimic receptor signaling in response to a specific antigen. While the RTK-based signalobodies enabled us to control cell growth and migration, further extension of applicability in another cell type would underlie the impact of the RTK-based signalobodies. In this study, we applied the scFv-c-Fms (S-Fms) signalobody in a murine myeloid progenitor cell line, FDC-P1. S-Fms transduced a fluorescein-conjugated BSA (BSA-FL)-dependent growth signal and activated downstream signaling molecules including MEK, ERK, Akt, and STAT3, which are major constituents of Ras/MAPK, PI3K/Akt, and JAK/STAT signaling pathways. In addition, S-Fms transduced a migration signal as demonstrated by the transwell-based migration assay. Direct real-time observation of the cells further confirmed that FDC/S-Fms cells underwent directional cell migration toward a positive gradient of BSA-FL. These results demonstrated the utility of the S-Fms signalobody for controlling growth and migration of myeloid cells. Further extension of our approach includes economical large-scale production of practically relevant blood cells as well as artificial control of cell migration for tissue regeneration and immune response.

Cell fate‐inducing CARs orthogonally control multiple signaling pathways

Nakajima

Nakabayashi

2022

Biotechnology Journal

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While chimeric antigen receptor (CAR) has been clinically applied in T cell‐mediated cancer therapy, CARs that combinatorially control general cell fates have yet to be practical. In this study, we aim to control multiple cell fates simultaneously by combinatorial activation of multiple cell fate‐inducing CARs (cfiCARs). We construct CARs that transduce two different signals using two antigen‐specific CARs. The CARs are co‐expressed pairwise on the cell surface to let the two antigens act alone or in combination, thus arbitrarily control multiple cell fates. We utilize signaling domains derived from natural receptors (gp130, receptor activator of nuclear factor κB [RANK], c‐Fms, and Fas) and tyrosine motif‐engineered artificial signaling domains that preferentially recruit target signaling molecules (Shc and STAT3) for inducing specific cell fates by the CARs. Consequently, the signaling molecules downstream of these receptors are activated orthogonally by the corresponding CAR‐specific antigens. Furthermore, two completely different cell fates (proliferation and death) are successfully controlled simultaneously or sequentially over time by adding the two antigens, demonstrating proliferation‐ and apoptosis‐inducing CARs (piCAR and aiCAR). Thus, cfiCARs will be applied for delineating the basic principle of cell fate control and improving the efficacy of cell therapy, which would significantly contribute to cell biology and future medicine.