Bh3 profiling identifies heterogeneous dependency on bcl-2 family members in multiple myeloma and predicts sensitivity to bh3 mimetics

Access through your institution Buy or subscribe Despite a significant improvement in the survival of patients with multiple myeloma (MM), the disease remains incurable and innovative

strategies are needed. Interactions among BCL-2 family proteins mainly determine cellular fate decision in response to drug therapy. Thus, anti-apoptotic members such as BCL-2, BCL-XL or

MCL-1 represent an attractive target for therapy.1 BH3 profiling is a functional assay that identifies the tumor cell’s addiction to these anti-apoptotic members.2 The oral BCL-2-specific

BH3 mimetic ABT-199 demonstrated very promising results in BCL-2-dependent malignancies such as chronic lymphoid leukemia and mantle cell lymphoma.3–5 To date, there has been no

determination of what proportion of MM cases are likely to be BCL-2 dependent. The present study demonstrated that MM is a heterogeneous disease with respect to BCL-2, BCL-XL or MCL-1

dependence. Moreover, identification by BH3 profiling of BCL-2 dependence predicted _in vitro_ sensitivity to BH3 mimetics. BH3 profiling is a unique, functional method to measure the

dependence to the anti-apoptotic proteins in live cancer cells. The present study demonstrates that MM is a heterogeneous disease regarding its dependence on anti-apoptotic proteins and

cannot be considered as monolithically BCL-2, BCL-XL or MCL-1 dependent. MCL-1 is expressed in MM cells at levels higher than normal plasma cells and its expression level has been shown to

affect clinical outcome.7, 8 Here, mitochondria from half of MM cell lines (4/8) and from almost one-third of primary samples (4/14) were found to be MCL-1 dependent. These results are

consistent with the requirement for MCL-1 for survival of many myeloma cells. BH3 profiling also identified a subset of BCL-2 and/or BCL-XL-dependent MM cells with relatively less dependence

on MCL-1 and correctly predicted sensitivity to the BH3 mimetics ABT-199 and 263. In our series, one MM cell line (MM1-S) and two primary samples (#7 and #11) were found to be sensitive to

ABT-263 but insensitive to ABT-199, that linked to their BCL-XL dependence determined using BH3 profiling. Even if these findings indicate that BCL-XL could be an attractive target for MM,

the BCL-XL-related platelet toxicity has impaired the clinical development of ABT-263.9 Previous studies identified MM cells sensitive to BH3 mimetics based on their BCL-2/MCL-1 mRNA

ratio10, 11 or interactions of BCL-XL and BCL-2 with BIM.12 From a practical point of view, BH3 profiling can be performed in just 3 h with fewer cells (5 × 104 plasma cells to determine

response to the BAD BH3 peptide). This consideration is of importance because bone marrow samples from MM patients usually contain a low percentage of plasma cells (the median percentage of

plasma cell in BM aspirate was 9% in our series). It has been previously reported that sensitivity to ABT-199 was restricted in MM patients with t(11;14) translocation.11 Here, among the

three ABT-199 sensitive samples, two were found to be t(11;14). By including previous data from Touzeau _et al._,11 sensitivity to ABT-199 from 29 different primary MM samples was analyzed.

Overall, seven samples (24%) were found to be sensitive to the drug (LD50<100 nM). Interestingly, six of these ABT-199 sensitive samples carried the t(11;14) translocation. Of note, the

remaining sensitive sample was found to be BCL-2 dependent according to BH3 profiling. Moreover, one sensitive patient sample was found to be negative for the t(11;14) translocation,

suggesting that BCL-2 dependence may exist beyond this cytogenetic subgroup. The positive and negative predictive values of BH3 profiling to predict ABT-199 sensitivity were 75% and 100%,

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support from NIH grants R01CA129974. CT was supported by the Fondation Française pour la Recherche contre le Myélome et les Gammapathies monoclonales (F.F.R.M.G.). AUTHOR CONTRIBUTIONS AL

and CT designed the research and wrote the manuscript. CT performed the experiments. CT, AL, JR, TNC analyzed the data. PR, KA, MA, SLG and PM provided myeloma cells. All the authors

critically reviewed the manuscript. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA C Touzeau, J Ryan, J

Guerriero, P Richardson, K Anderson & A Letai * Department of Hematology, University hospital, Nantes, France C Touzeau, P Moreau & S Le Gouill * INSERM UMR892, CNRS UMR6299,

University of Nantes, Nantes, France C Touzeau, P Moreau, S Le Gouill & M Amiot * Conway Institute of Biomolecular and Biomedical Research, Dublin, Ireland T N Chonghaile Authors * C

Touzeau View author publications You can also search for this author inPubMed Google Scholar * J Ryan View author publications You can also search for this author inPubMed Google Scholar * J

Guerriero View author publications You can also search for this author inPubMed Google Scholar * P Moreau View author publications You can also search for this author inPubMed Google

Scholar * T N Chonghaile View author publications You can also search for this author inPubMed Google Scholar * S Le Gouill View author publications You can also search for this author

inPubMed Google Scholar * P Richardson View author publications You can also search for this author inPubMed Google Scholar * K Anderson View author publications You can also search for this

author inPubMed Google Scholar * M Amiot View author publications You can also search for this author inPubMed Google Scholar * A Letai View author publications You can also search for this

author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to A Letai. ETHICS DECLARATIONS COMPETING INTERESTS AL has been a paid advisor to AbbVie. AL’s laboratory has received

sponsorship for research with AbbVie. ADDITIONAL INFORMATION Supplementary Information accompanies this paper on the Leukemia website SUPPLEMENTARY INFORMATION SUPPLEMENTARY TABLE (PPT 151

KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Touzeau, C., Ryan, J., Guerriero, J. _et al._ BH3 profiling identifies heterogeneous dependency on

Bcl-2 family members in multiple myeloma and predicts sensitivity to BH3 mimetics. _Leukemia_ 30, 761–764 (2016). https://doi.org/10.1038/leu.2015.184 Download citation * Published: 15 July

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