Abstract
Malignant peripheral nerve sheath tumors (MPNST) are highly aggressive soft tissue sarcomas with limited treatment options, and novel effective therapeutic strategies are desperately needed. We observe anti-proliferative efficacy of genetic depletion or pharmacological inhibition using the clinically available SHP2 inhibitor (SHP2i) TNO155. Our studies into the signaling response to SHP2i reveal that resistance to TNO155 is partially mediated by reduced RB function, and we therefore test the addition of a CDK4/6 inhibitor (CDK4/6i) to enhance RB activity and improve TNO155 efficacy. In combination, TNO155 attenuates the adaptive response to CDK4/6i, potentiates its anti-proliferative effects, and converges on enhancement of RB activity, with greater suppression of cell cycle and inhibitor-of-apoptosis proteins, leading to deeper and more durable anti-tumor activity in in vitro and in vivo patient-derived models of MPNST, relative to either single agent. Overall, our study provides timely evidence to support the clinical advancement of this combination strategy in patients with MPNST and other tumors driven by loss of NF1.
Competing Interest Statement
JW, SEM and CAP have a pending patent related to this study. The authors have no additional financial interests.
Footnotes
Summary: Effective treatment approaches are an unmet clinical need for MPNST. This work proves the dependency of NF1-MPNST on SHP2/PTPN11, and demonstrates the anti-tumor efficacy and well-tolerability of SHP2i plus CDK4/6i, through enhancement of RB function, in patient-derived models of MPNST.
Abbreviations
- 3D
- Three dimensional
- 60M
- 60 million
- 7-AAD
- 7-aminoactinomycin D
- adj
- adjusted
- AKT
- Ak strain transforming
- ANOVA
- Analysis of variance
- AS
- Anti-sense
- AUC
- Area under the curve
- AURKA
- Aurora kinase A
- BAF
- B-allele frequency
- BAM
- Binary Alignment Map
- BCA
- Bicinchoninic acid
- BD
- Becton, Dickinson and Company
- BED
- Browser Extensible Data
- BIRC5
- Baculoviral IAP repeat containing 5
- BQSR
- Base quality score recalibration
- BWA-MEM
- Burrows-Wheeler Alignment Tool - Maximal Exact Match
- °C
- Degree Celsius
- CC1 buffer
- Cell Conditioning buffer
- CCK-8
- Cell Counting Kit-8
- CCNA2
- Cyclin A2
- CDK
- Cyclin dependent kinase
- CDK4/6i
- CDK4/6 inhibitor
- CDKN2A
- Cyclin dependent kinase inhibitor 2A
- CLSPN
- Claspin
- cm
- Centimeter
- ctrl
- Control
- DAB
- 3,3′-Diaminobenzidine
- DEG
- Differentially expressed genes
- DMSO
- Dimethyl sulfoxide
- Dox
- Doxycycline
- DPBS
- Dulbecco’s phosphate-buffered saline
- ECL
- Enhanced chemiluminescence
- EED
- Embryonic Ectoderm Development
- ERBB3
- Erb-b2 receptor tyrosine kinase 3
- ERK
- Extracellular signal-regulated kinase
- F
- Forward
- FACS
- Fluorescence-Activated Cell Sorting
- FBS
- Fetal bovine serum
- FOXM1
- Forkhead box M1
- G
- Germline
- GAP
- GTPase-activating protein
- GATK
- Genome Analysis Toolkit
- gDNA
- genomic deoxyribonucleic acid
- GEF
- Guanine nucleotide exchange factors
- GFP
- Green fluorescent protein
- ggplot2
- Grammar of graphics plot 2
- GnomAD
- Genome Aggregation Database
- GRCh38
- Genome Reference Consortium Human Build 38
- GRD
- GAP related domain
- GTP
- Guanosine triphosphate
- HEK
- Human embryonic kidney
- HPV
- Human papillomavirus
- HRP
- Horseradish peroxidase
- IACUC
- Institutional Animal Care and Use Committee
- IAP
- The inhibitor of apoptosis proteins
- IC50
- The half maximal inhibitory concentration
- IGF-IR
- Insulin like growth factor 1 receptor
- IHC
- Immunohistochemistry
- indels
- Insertions and deletions
- JH
- Johns Hopkins
- KEGG
- Kyoto Encyclopedia of Genes and Genomes
- KIR3DL2
- Killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 2
- L
- Liter
- LFC
- Logarithm fold change
- LKO
- Lentiviral knockout
- LOF
- Loss of function
- Log
- logarithm
- MAF
- Minor allele frequency
- MEK
- Mitogen-activated protein kinase kinase, MAP2K
- mg
- Milligram
- ml
- Milliliter
- MN
- University of Minnesota
- MPNST
- Malignant peripheral nerve sheath tumors
- MSCV
- Murine Stem Cell Virus
- NBF
- Neutral buffered formalin
- NCT
- National clinical trials
- NF1
- Neurofibromatosis Type 1
- NF1
- Neurofibromin 1
- NFRI
- NF Research Initiative
- ng
- nanogram
- NIBR
- Novartis Institute for Biomedical Research
- NIH
- National Institutes of Health
- NRG
- NOD Rag gamma
- ns
- not significant
- NTRK2
- Neurotrophic Receptor Tyrosine Kinase 2
- Par
- Parental
- PBS
- Phosphate-buffered saline
- PCR
- Polymerase chain reaction
- PD
- Pharmacodynamic
- PD
- Pull down
- Portable document format
- PDX
- Patient-derived xenograft
- PE
- Paired-end
- pH
- Potential of hydrogen
- PI
- Propidium iodide
- PIK3CA
- The phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha
- PLK-1
- Polo-like kinase 1
- pNF
- Plexiform neurofibromas
- PRC2
- Polycomb repressive complex 2
- PS
- Partially sensitive
- PTPN11
- Protein tyrosine phosphatase non-receptor type 11
- puro
- Puromycin
- PVDF
- Polyvinylidene fluoride
- R
- Reverse
- RABL6A
- Rab-like protein 6A
- RAF
- Rapidly accelerated fibrosarcoma
- RAS
- Rat sarcoma
- RB
- Retinoblastoma 1
- RBC
- Red blood cells
- ref
- Reference
- Res
- Resistant
- RIPA
- Radio-Immunoprecipitation Assay
- RNase
- Ribonuclease
- RNAseq
- RNA sequencing
- rRNA
- Ribosomal ribonucleic acid
- RTK
- Receptor tyrosine kinases
- rtTA
- Reverse tetracycline-controlled trans-activator
- S
- Somatic
- S
- Sense
- SD
- Standard deviation
- SDS-PAGE
- Sodium dodecyl-sulfate polyacrylamide gel electrophoresis
- SEM
- Standard error of the mean
- SHP2
- Src homology-2 domain-containing protein tyrosine phosphatase-2
- SHP2i
- SHP2 inhibitor
- shRNA
- Short/small hairpin RNA
- SKCCC
- Sidney Kimmel Comprehensive Cancer Center
- SNV
- Single nucleotide variant
- STR
- Short-tandem repeat
- SUZ12
- Suppressor Of Zeste 12 Protein Homolog
- TAZ
- Tafazzin
- Tet
- Tetracycline
- TP53
- Tumor protein P53
- tram
- trametinib
- TSC2
- Tuberous Sclerosis Complex 2
- μg
- Microgram
- μm
- Micrometer
- μmol
- Micromole
- v.
- versus
- VEP
- Variant Effect Predictor
- VS
- Very sensitive
- WCL
- whole cell lysate
- WES
- Whole exome sequencing
- WNT2B
- Wnt family member 2B
- WT
- Wild-type
- WU
- University of Washington
- YAP
- Yes-associated protein
