Generation of Non-Small Cell Lung Cancer Patient-Derived Xenografts to Study Intratumor Heterogeneity
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Patient Recruitment
2.2. PDX Generation
2.3. Histopathological Examination
2.4. DNA/RNA Extraction
2.5. Library Preparation and Sequencing
2.6. Primary and PDX Tumor Comparison
2.7. CTC Enrichment for Molecular Analysis
2.8. Gene Expression Analysis of CTCs by RT-qPCR
2.9. Statistical Analysis
3. Results
3.1. Establishment of NSCLC PDXs
3.2. Clinical Characteristics and Grafting Efficiency
3.3. Correlation between the Presence of CTCs and Grafting
3.4. Molecular Comparison of PDX and Primary Tumors
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Patient Characteristics | PDX YES (n = 31) | PDX NO (n = 21) | p-Value |
---|---|---|---|
Categorical Variables | n (%) | n (%) | |
Gender | 0.141 a | ||
Male | 23 (74.2%) | 11 (52.4%) | |
Female | 8 (25.8%) | 10 (47.6%) | |
Age | 0.090 a | ||
<68y b | 10 (34.5%) | 12 (60.0%) | |
≥68y | 19 (65.5%) | 8 (40.0%) | |
Smoking status | 0.497 c | ||
Current | 19 (61.3%) | 10 (47.6%) | |
Previous | 6 (19.4%) | 7 (33.3%) | |
Never | 6 (19.4%) | 4 (19.0%) | |
Histological type | 0.100 c | ||
Adenocarcinoma | 10 (32.3%) | 13 (61.9%) | |
Squamous | 17 (54.8%) | 7 (33.3%) | |
Large cell or Sarcomatous | 4 (12.9%) | 1 (4.8%) | |
Clinical stage | 0.040 c | ||
I | 13 (41.9%) | 10 (47.6%) | |
II | 6 (19.4%) | 9 (42.9%) | |
IIIA | 12 (38.7%) | 2 (9.5%) | |
Tumor stage | 0.038 c | ||
pT1 | 5 (16.1%) | 9 (42.9%) | |
pT2 | 15 (48.4%) | 7 (33.3%) | |
pT3 | 5 (16.1%) | 5 (23.8%) | |
pT4 | 6 (19.4%) | 0 (0%) | |
Continuous variables | Median (min–max) | Median (min–max) | |
Tumor volume (cm3) | 0.070 d | ||
33.7 (0.48–572.0) | 14.7 (2.14–245.0) | ||
Age (years) | 0.030 d | ||
71 (56–81) | 64 (40–75) |
PDX | Total | p Value a | |||
---|---|---|---|---|---|
No (−) | Yes (+) | ||||
KRT18 | Neg | 21 | 19 | 37 | p = 0.009 |
Pos | 0 | 7 | 7 | ||
Total | 21 | 26 | 47 | ||
KRT19 | Neg | 16 | 19 | 34 | p = 1.000 |
Pos | 5 | 7 | 12 | ||
Total | 21 | 26 | 47 | ||
KRT8 | Neg | 17 | 22 | 39 | p = 1.000 |
Pos | 4 | 4 | 8 | ||
Total | 21 | 26 | 47 | ||
Epithelial markers | Neg | 14 | 14 | 28 | p = 0.377 |
Pos | 7 | 12 | 19 | ||
Total | 21 | 26 | 47 |
PDX | Total | p Value a | |||
---|---|---|---|---|---|
No (−) | Yes (+) | ||||
VIM | Neg | 11 | 9 | 20 | p = 0.231 |
Pos | 10 | 17 | 27 | ||
Total | 21 | 26 | 47 | ||
TWIST1 | Neg | 18 | 17 | 35 | p = 0.179 |
Pos | 3 | 9 | 12 | ||
Total | 21 | 26 | 47 | ||
EMT markers (VIM & TWIST1) | Neg | 8 | 5 | 13 | p = 0.107 |
Pos | 13 | 21 | 34 | ||
Total | 21 | 26 | 47 |
CODE | TYPE | GENETIC ALTERATIONS | TUMOR CELLULARITY (%) | VAFPRIM.TUMOR | VAFPDX |
---|---|---|---|---|---|
105 | LUSC | TP53:c.469G>T:p.V157F KMT2C:c.918T>G:p.Y306* | 90 | 0.057 0.059 | 1 0.062 |
110 | LUSC | TP53:c.413C>T:p.A138V ALK:c.2712T>A:p.H904Q | 85 | 0.432 0.36 | 0.952 0.191 |
112 | PLEIO/LUAD | CTNNB1:c.98C>T:p.S33F TP53:c.811G>T:p.E271* STK11:c.487G>T:p.G163C KDR:c.2761delinsAT:p.F921Ifs*13 KDR:c.2757C>G:p.C919W | 80 | 0.49 0.436 0.356 0.316 0.317 | 0.868 0.93 0.895 0.83 0.831 |
302 | LUSC | KIT:2508G>T:p.M836I EGFR:c.1150A>T:p.T384S, MEN1:c.184A>G:p.T62A, MET:c.1241A>G:p.D414G | 70 | 0.28 0.392 0.325 0.64 | 0.973 0.993 0.644 0.995 |
512 | LUSC | TP53:c.845G>C:p.R282P CDKN2A:c.316_317T:p.V106Cfs*39 PDGFRA:c.1102G>C:p.E368Q | 75 | 0.336 0.375 0.4 | 0.999 0.998 0.995 |
517 | LUSC | RB1:c.852delinsTA:p.I285Nfs*2 | 75 | 0.249 | 0.998 |
518 | LUSC | TP53:c.659A>G:p.Y220C | 80 | 0.304 | 0.996 |
519 | LCNEC | RB1:c.1953_1954T:p.V654Cfs*3 | 80 | 0.732 | 0.998 |
531 | LUSC | TP53:c.747G>T:p.R249S CDKN2A:c.262G>T:p.E88* PIK3CA:c.1633G>A:p.E545K | 80 | 0.574 0.416 0.251 | 0.998 0.991 0.337 |
546 | LUAD | KRAS:c.35G>A:p.G12D ATM:c.8851G>A:p.V2951I | 85 | 0.342 0.371 | 0.296 0.997 |
551 | LUSC | MTOR c.1333A>G:p.R445G CDKN2A:c.61G>C:p.A21P | 65 | 0.686 0.295 | 0.994 0.966 |
568 | LUAD | TP53:c.880G>T:p.E294* KRAS:c.34G>T:p.G12C | 65 | 0.09 0.16 | 0.63 0.62 |
569 | LUSC | NF1:c.4662-1G>T TP53:c.994-2A>T; SMAD4:c.1326G>T:p.Q442H | 80 | 0.301 0.274 0.145 | 0.984 0.991 0.953 |
571 | LUSC | CHEK2: c.1114C>T:p.Q372* TP53:c.994-2A>T BRCA2:c.3668A>G:p.H1223R NOTCH1:c.2058_2059T:p.C687Afs*84 | 75 | 0.294 0.287 0.59 0.279 | 0.362 0.952 0.959 0.951 |
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Kanaki, Z.; Voutsina, A.; Markou, A.; Pateras, I.S.; Potaris, K.; Avgeris, M.; Makrythanasis, P.; Athanasiadis, E.I.; Vamvakaris, I.; Patsea, E.; et al. Generation of Non-Small Cell Lung Cancer Patient-Derived Xenografts to Study Intratumor Heterogeneity. Cancers 2021, 13, 2446. https://doi.org/10.3390/cancers13102446
Kanaki Z, Voutsina A, Markou A, Pateras IS, Potaris K, Avgeris M, Makrythanasis P, Athanasiadis EI, Vamvakaris I, Patsea E, et al. Generation of Non-Small Cell Lung Cancer Patient-Derived Xenografts to Study Intratumor Heterogeneity. Cancers. 2021; 13(10):2446. https://doi.org/10.3390/cancers13102446
Chicago/Turabian StyleKanaki, Zoi, Alexandra Voutsina, Athina Markou, Ioannis S. Pateras, Konstantinos Potaris, Margaritis Avgeris, Periklis Makrythanasis, Emmanouil I. Athanasiadis, Ioannis Vamvakaris, Eleni Patsea, and et al. 2021. "Generation of Non-Small Cell Lung Cancer Patient-Derived Xenografts to Study Intratumor Heterogeneity" Cancers 13, no. 10: 2446. https://doi.org/10.3390/cancers13102446