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siRNA targeting the κ light chain constant region: preclinical testing of an approach to nonfibrillar and fibrillar light chain deposition diseases

Gene Therapy volume 23, pages 727–733 (2016)Cite this article

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Abstract

Treatment of light chain (LC) deposition diseases both nonfibrillar and fibrillar is aimed at eliminating LC production but success is limited. We report on the testing of an small interfering RNA pool targeting the κ LC constant region mRNA (si[IGKC]) designed for use against all κ plasma cell clones. To test for changes in κ LC message and protein production we used real-time PCR, immunoblot, intracellular mean fluorescence intensity and κ LC secretion by enzyme-linked immunosorbent assay. In vitro we employed 4 human cell lines that make κ LCs and 20 specimens of CD138-selected marrow plasma cells from patients with κ plasma cell diseases. In vivo, we used a murine flank plasmacytoma xenograft model. In vitro and in vivo, there were significant reductions in message and protein production by all modalities in all cell types despite diversity in variable region sequence. In addition, in clones producing intact immunoglobulin, caspase 3/7 activity with si[IGKC] was significantly increased compared with clones producing κ LC only, consistent with the triggering of a terminal unfolded protein response by excess unpaired heavy chains. In conclusion, si[IGKC] can significantly reduce κ LC production by κ plasma cells. Further preclinical development is needed to optimize delivery.

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Acknowledgements

We thank the Division of Hematology-Oncology and Departments of Medicine and Pathology at Tufts for their continued support. We also acknowledge the continued support by the Amyloidosis and Myeloma Research Fund at Tufts, the Cam Neely and John Davis Myeloma Research Fund, the John C Davis Program for Myeloma and Amyloid at Tufts, the Sidewater Family Fund, the Lavonne Horowitz Trust, the Werner and Elaine Dannheiser Fund for Research on the Biology of Aging of the Lymphoma Foundation, Davis and Barbara Levine in memoriam and especially the Demarest Lloyd Jr Foundation for their continued commitment to ‘shutting down the factory’ in AL.

Author contributions

XM and PZ designed and performed experiments, analyzed data and wrote the manuscript; SWW, MW and CC obtained patient specimens and wrote the manuscript; RLC designed the research, analyzed data and wrote the manuscript.

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Author notes

  1. X Ma and P Zhou: These authors contributed equally to this work.

Authors and Affiliations

  1. Tufts Medical Center, Boston, MA, USA

    X Ma, S W Wong, M Warner & R L Comenzo

  2. Tufts Medical Center, Tufts University School of Medicine, Boston, MA, USA

    P Zhou

  3. Taussig Cancer Institute of Cleveland Clinic, Cleveland Clinic Florida, Weston, FL, USA

    C Chaulagain

Authors
  1. X Ma
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  2. P Zhou
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Corresponding author

Correspondence to R L Comenzo.

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Competing interests

Drs Comenzo, Ma and Zhou have a patent pending on the siRNA sequences used to knock down expression of the κ light chain constant region. Dr Comenzo has served as a consultant or advisor to Takeda Millennium and Janssen, and has received clinical research funding from Takeda Millennium, Janssen, Teva and Prothena Biotech. Drs Wong and Chaulagain and Ms Warner declare no conflict of interest.

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Ma, X., Zhou, P., Wong, S. et al. siRNA targeting the κ light chain constant region: preclinical testing of an approach to nonfibrillar and fibrillar light chain deposition diseases. Gene Ther 23, 727–733 (2016). https://doi.org/10.1038/gt.2016.50

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