CoChat
Sign Up
Cited by 15RNA Interference and Gene DeliveryPancreatic and Hepatic Oncology ResearchDNA and Nucleic Acid Chemistry

Inhibiting mutant KRAS G12D gene expression using novel peptide nucleic acid‑based antisense: A potential new drug candidate for pancreatic cancer

Open access
Shai, Ayelet, Galouk, Evleen, Miari, Reem, Tareef, Hala, Sammar, Marei, Zeidan, Mouhammad, Rayan, Anwar, Falah, Mizied·2022·Oncology Letters
DOI 10.3892/ol.2022.13250PMC PMC8895471
Open this paper in CoChat — free accountOpen PDF

Abstract

KRAS mutations, which are the main cause of the pathogenesis of lethal pancreatic adenocarcinomas, impair the functioning of the GTPase subunit, thus rendering it constitutively active and signaling intracellular pathways that end with cell transformation. In the present study, the AsPC-1 cell line, which has a G12D-mutated KRAS gene sequence, was utilized as a cellular model to test peptide nucleic acid-based antisense technology. The use of peptide nucleic acids (PNAs) that are built to exhibit improved hybridization specificity and have an affinity for complementary RNA and DNA sequences, as well as a simple chemical structure and high biological stability that affords resistance to nucleases and proteases, enabled targeting of the KRAS-mutated gene to inhibit its expression at the translation level. Because PNA-based antisense molecules should be capable of binding to KRAS mRNA sequences, PNAs were utilized to target the mRNA of the mutated KRAS gene, a strategy that could lead to the development of a novel drug for pancreatic cancer. Moreover, it was demonstrated that introducing new PNA to cells inhibited the growth of cancer cells and induced apoptotic death and, notably, that it can inhibit G12D-mutated KRAS gene expression, as demonstrated by RT-PCR and western blotting. Altogether, these data strongly suggest that the use of PNA-based antisense agents is an attractive therapeutic approach to treating KRAS-driven cancers and may lead to the development of novel drugs that target the expression of other mutated genes.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.3892/ol.2022.13250

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • This allows PNAs to outcompete the antisense strand to selectively hybridize with the mutant region of the genome, thus obstructing transcriptional machinery and causing reduced expression.

    “This allows PNAs to outcompete the antisense strand to selectively hybridize with the mutant region of the genome, thus obstructing transcriptional machinery and causing reduced expression [71].”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [71]

    The Introduction says PNAs bind complementary sequences with high specificity, and the Results state that these PNAs were designed to target mutant KRAS mRNA. The Discussion reports reduced mutant KRAS translation and protein. The paper does not show PNAs outcompeting an antisense strand or obstructing transcriptional machinery at the genomic region; those mechanisms are not supported for this experiment. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • The avidity of PNAs for a single strand of matching DNA is higher than DNA itself.

    “The avidity of PNAs for a single strand of matching DNA is higher than DNA itself.”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [71]

  • Engineered peptide nucleic acids (PNAs) have protein-like backbone structures bound to DNA-like bases.

    “Engineered peptide nucleic acids (PNAs) have protein-like backbone structures bound to DNA-like bases.”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [71]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

No citation lists for this paper

No citation lists for this paper yet — checked again daily.

References 53 · Cited by 15

Research works printed but not linked: 47

  1. [1]Prior IA Lewis PD Mattos C A comprehensive survey of Ras mutations in cancer Cancer Res 72 2457 2467 2012 10.1158/0008-5472.CAN-11-2612 22589270 PMC3354961 · DOI · Paper page
  2. [2]Fernández-Medarde A Santos E Ras in cancer and developmental diseases Genes Cancer 2 344 358 2011 10.1177/1947601911411084 21779504 PMC3128640 · DOI
  3. [4]Tsiatis AC Norris-Kirby A Rich RG Hafez MJ Gocke CD Eshleman JR Murphy KM Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: Diagnostic and clinical implications J Mol Diagn 12 425 432 2010 10.2353/jmoldx.2010.090188 20431034 PMC2893626 · DOI
  4. [5]Pylayeva-Gupta Y Grabocka E Bar-Sagi D RAS oncogenes: Weaving a tumorigenic web Nat Rev Cancer 11 761 774 2011 10.1038/nrc3106 21993244 PMC3632399 · DOI · Paper page
  5. [6]Singh H Longo DL Chabner BA Improving prospects for targeting RAS J Clin Oncol 33 3650 3659 2015 10.1200/JCO.2015.62.1052 26371146 · DOI
  6. [8]Buscail L Bournet B Cordelier P Role of oncogenic KRAS in the diagnosis, prognosis and treatment of pancreatic cancer Nat Rev Gastroenterol Hepatol 17 153 168 2020 10.1038/s41575-019-0245-4 32005945 · DOI · Paper page
  7. [10]Scheffzek K Ahmadian MR Kabsch W Wiesmüller L Lautwein A Schmitz F Wittinghofer A The Ras-RasGAP complex: Structural basis for GTPase activation and its loss in oncogenic Ras mutants Science 277 333 338 1997 10.1126/science.277.5324.333 9219684 · DOI · Paper page
  8. [11]Bos JL Rehmann H Wittinghofer A GEFs and GAPs: Critical elements in the control of small G proteins Cell 129 865 877 2007 10.1016/j.cell.2007.05.018 17540168 · DOI · Paper page
  9. [13]Liu F Yang X Geng M Huang M Targeting ERK, an Achilles' Heel of the MAPK pathway, in cancer therapy Acta Pharm Sin B 8 552 562 2018 10.1016/j.apsb.2018.01.008 30109180 PMC6089851 · DOI
  10. [14]Vallejo A Perurena N Guruceaga E Mazur PK Martinez-Canarias S Zandueta C Valencia K Arricibita A Gwinn D Sayles LC An integrative approach unveils FOSL1 as an oncogene vulnerability in KRAS-driven lung and pancreatic cancer Nat Commun 8 14294 2017 10.1038/ncomms14294 28220783 PMC5321758 · DOI
  11. [15]Ryan MB Corcoran RB Therapeutic strategies to target RAS-mutant cancers Nat Rev Clin Oncol 15 709 720 2018 10.1038/s41571-018-0105-0 30275515 · DOI · Paper page
  12. [16]Takashima A Faller DV Targeting the RAS oncogene Expert Opin Ther Targets 17 507 531 2013 10.1517/14728222.2013.764990 23360111 PMC3804031 · DOI
  13. [17]Zorde Khvalevsky E Gabai R Rachmut IH Horwitz E Brunschwig Z Orbach A Shemi A Golan T Domb AJ Yavin E Mutant KRAS is a druggable target for pancreatic cancer Proc Natl Acad Sci USA 110 20723 20728 2013 10.1073/pnas.1314307110 24297898 PMC3870687 · DOI · Paper page
  14. [18]Ross SJ Revenko AS Hanson LL Ellston R Staniszewska A Whalley N Pandey SK Revill M Rooney C Buckett LK Targeting KRAS-dependent tumors with AZD4785, a high-affinity therapeutic antisense oligonucleotide inhibitor of KRAS Sci Transl Med 9 eaal5253 2017 10.1126/scitranslmed.aal5253 28615361 · DOI · Paper page
  15. [19]Marin VL Roy S Armitage BA Recent advances in the development of peptide nucleic acid as a gene-targeted drug Expert Opin Biol Ther 4 337 348 2004 10.1517/14712598.4.3.337 15006728 · DOI
  16. [20]Gambari R Peptide-nucleic acids (PNAs): A tool for the development of gene expression modifiers Curr Pharm Des 7 1839 1862 2001 10.2174/1381612013397087 11562312 · DOI
  17. [21]Nielsen PE Egholm M Berg RH Buchardt O Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide Science 254 1497 1500 1991 10.1126/science.1962210 1962210 · DOI · Paper page
  18. [22]Rajasekaran P Alexander JC Seleem MN Jain N Sriranganathan N Wattam AR Setubal JC Boyle SM Peptide nucleic acids inhibit growth of Brucella suis in pure culture and in infected murine macrophages Int J Antimicrob Agents 41 358 362 2013 10.1016/j.ijantimicag.2012.11.017 23305655 PMC3834731 · DOI
  19. [23]Abdel-Aziz M Yamasaki T Otsuka M Synthesis and hybridization property of novel 2′,5′-isoDNA mimic chiral peptide nucleic acids Bioorg Med Chem Lett 13 1041 1043 2003 10.1016/S0960-894X(03)00075-1 12643907 · DOI
  20. [24]Koppelhus U Nielsen PE Cellular delivery of peptide nucleic acid (PNA) Adv Drug Deliv Rev 55 267 280 2003 10.1016/S0169-409X(02)00182-5 12564980 · DOI
  21. [25]Dean DA Peptide nucleic acids: Versatile tools for gene therapy strategies Adv Drug Deliv Rev 44 81 95 2000 10.1016/S0169-409X(00)00087-9 11072107 PMC4403640 · DOI
  22. [26]De Cola C Manicardi A Corradini R Izzo I De Riccardis F Carboxyalkyl peptoid PNAs: Synthesis and hybridization properties Tetrahedron 68 499 506 2012 10.1016/j.tet.2011.11.017 · DOI
  23. [27]Narenji H Gholizadeh P Aghazadeh M Rezaee MA Asgharzadeh M Kafil HS Peptide nucleic acids (PNAs): Currently potential bactericidal agents Biomed Pharmacother 93 580 588 2017 10.1016/j.biopha.2017.06.092 28686972 · DOI
  24. [28]Wang G Xu XS Peptide nucleic acid (PNA) binding-mediated gene regulation Cell Res 14 111 116 2004 10.1038/sj.cr.7290209 15115611 · DOI
  25. [29]Quijano E Bahal R Ricciardi A Saltzman WM Glazer PM Therapeutic peptide nucleic acids: Principles, limitations, and opportunities Yale J Biol Med 90 583 598 2017 29259523 PMC5733847
  26. [30]Zhao XL Chen BC Han JC Wei L Pan XB Delivery of cell-penetrating peptide-peptide nucleic acid conjugates by assembly on an oligonucleotide scaffold Sci Rep 5 17640 2015 10.1038/srep17640 26612536 PMC4661726 · DOI
  27. [31]Lucarelli F Marrazza G Turner AP Mascini M Carbon and gold electrodes as electrochemical transducers for DNA hybridisation sensors Biosens Bioelectron 19 515 530 2004 10.1016/S0956-5663(03)00256-2 14683635 · DOI
  28. [32]Barluenga S Winssinger N PNA as a biosupramolecular tag for programmable assemblies and reactions Acc Chem Res 48 1319 1331 2015 10.1021/acs.accounts.5b00109 25947113 · DOI · Paper page
  29. [33]Brandén LJ Mohamed AJ Smith CI A peptide nucleic acid-nuclear localization signal fusion that mediates nuclear transport of DNA Nat Biotechnol 17 784 787 1999 10.1038/11726 10429244 · DOI
  30. [34]Liang KW Hoffman EP Huang L Targeted delivery of plasmid DNA to myogenic cells via transferrin-conjugated peptide nucleic acid Mol Ther 1 236 243 2000 10.1006/mthe.2000.0043 10933939 · DOI
  31. [35]Chiarantini L Cerasi A Fraternale A Millo E Benatti U Sparnacci K Laus M Ballestri M Tondelli L Comparison of novel delivery systems for antisense peptide nucleic acids J Control Release 109 24 36 2005 10.1016/j.jconrel.2005.09.013 16290244 · DOI
  32. [36]Montazersaheb S Hejazi MS Nozad Charoudeh H Potential of peptide nucleic acids in future therapeutic applications Adv Pharm Bull 8 551 563 2018 10.15171/apb.2018.064 30607328 PMC6311635 · DOI
  33. [37]Elbashir SM Harborth J Lendeckel W Yalcin A Weber K Tuschl T Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells Nature 411 494 498 2001 10.1038/35078107 11373684 · DOI · Paper page
  34. [38]Fire A Xu S Montgomery MK Kostas SA Driver SE Mello CC Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans Nature 391 806 811 1998 10.1038/35888 9486653 · DOI · Paper page
  35. [39]Kennerdell JR Carthew RW Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway Cell 95 1017 1026 1998 10.1016/S0092-8674(00)81725-0 9875855 · DOI
  36. [42]Komiyama M Aiba Y Ishizuka T Sumaoka J Solid-phase synthesis of pseudo-complementary peptide nucleic acids Nat Protoc 3 646 654 2008 10.1038/nprot.2008.6 18388947 · DOI
  37. [44]Ostrem JM Peters U Sos ML Wells JA Shokat KM K-Ras(G12C) inhibitors allosterically control GTP affinity and effector interactions Nature 503 548 551 2013 10.1038/nature12796 24256730 PMC4274051 · DOI · Paper page
  38. [46]Xu CF Wang J Delivery systems for siRNA drug development in cancer therapy Asian J Pharm Sci 10 1 12 2015 10.1016/j.ajps.2014.08.011 · DOI
  39. [47]Llovera L Berthold P Nielsen PE Shiraishi T Cell number and transfection volume dependent peptide nucleic acid antisense activity by cationic delivery methods Artif DNA PNA XNA 3 22 27 2012 10.4161/adna.19906 22679530 PMC3368813 · DOI
  40. [48]Hartwell LH Weinert TA Checkpoints: Controls that ensure the order of cell cycle events Science 246 629 634 1989 10.1126/science.2683079 2683079 · DOI
  41. [49]Malumbres M Barbacid M Cell cycle, CDKs and cancer: A changing paradigm Nat Rev Cancer 9 153 166 2009 10.1038/nrc2602 19238148 · DOI · Paper page
  42. [50]Chinnasamy S Zameer F Muthuchelian K Molecular and biological mechanisms of apoptosis and its detection techniques J Oncol Sci 6 49 64 2020 10.37047/jos.2020-73477 · DOI
  43. [51]Evan GI Vousden KH Proliferation, cell cycle and apoptosis in cancer Nature 411 342 348 2001 10.1038/35077213 11357141 · DOI
  44. [52]Wong RS Apoptosis in cancer: From pathogenesis to treatment J Exp Clin Cancer Res 30 87 2011 10.1186/1756-9966-30-87 21943236 PMC3197541 · DOI · Paper page
  45. [53]Rachagani S Senapati S Chakraborty S Ponnusamy MP Kumar S Smith LM Jain M Batra SK Activated KrasG¹2D is associated with invasion and metastasis of pancreatic cancer cells through inhibition of E-cadherin Br J Cancer 104 1038 1048 2011 10.1038/bjc.2011.31 21364589 PMC3065271 · DOI
  46. [54]Smakman N Veenendaal LM van Diest P Bos R Offringa R Borel Rinkes IH Kranenburg O Dual effect of Kras(D12) knockdown on tumorigenesis: Increased immune-mediated tumor clearance and abrogation of tumor malignancy Oncogene 24 8338 8342 2005 10.1038/sj.onc.1208995 16091732 · DOI
  47. [55]Gupta A Bahal R Gupta M Glazer PM Saltzman WM Nanotechnology for delivery of peptide nucleic acids (PNAs) J Control Release 240 302 311 2016 10.1016/j.jconrel.2016.01.005 26776051 PMC5016210 · DOI

Authors
Shai, A., Galouk, E., Miari, R., Tareef, H., Sammar, M., Zeidan, M., Rayan, A., & Falah, M.
Affiliations
  • Western Galilee Hospital (IL)
  • Braude College of Engineering Karmiel (IL)
  • Molecular Genetics and Virology Laboratory, Al‑Qasemi Center of Research Excellence, Baka EL‑Garbiah 30100, Israel
  • Al-Qasemi Academic College of Education (IL)
Corresponding author
  • Rayan, Anwar
  • Falah, Mizied
ORCID
Venue
Oncology Letters
Publisher
Spandidos Publications
Published
2022-02-21
Volume
23
Issue
4
Pages
130-130
ISSN
  • 1792-1074 (ISSN-L)
  • 1792-1082
DOI
10.3892/ol.2022.13250
PMC
PMC8895471
OpenAlex
W4213306578
Cited by
15
References
53
Citations by year
2026: 4 · 2025: 3 · 2024: 1 · 2023: 6 · 2022: 1
FWCI
0.90
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Oncology Letters (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Health Sciences › Medicine › Oncology › Pancreatic and Hepatic Oncology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords
KRAS G12D mutation, peptide nucleic acid, antisense oligonucleotides, pancreatic ductal adenocarcinoma, AsPC-1 cells, protein synthesis inhibition, apoptosis, anticancer drug development, KRAS-driven cancers

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review