Limits...
Presence of cancer-associated mutations in exhaled breath condensates of healthy individuals by next generation sequencing

View Article: PubMed Central - PubMed

ABSTRACT

Exhaled breath condensate (EBC) is a non-invasive source that can be used for studying different genetic alterations occurring in lung tissue. However, the low yield of DNA available from EBC has hampered the more detailed mutation analysis by conventional methods. We applied the more sensitive amplicon-based next generation sequencing (NGS) to identify cancer related mutations in DNA isolated from EBC. In order to apply any method for the purpose of mutation screening in cancer patients, it is important to clarify the incidence of these mutations in healthy individuals. Therefore, we studied mutations in hotspot regions of 22 cancer genes of 20 healthy, mainly non-smoker individuals, using AmpliSeq colon and lung cancer panel and sequenced on Ion PGM.

In 15 individuals, we detected 35 missense mutations in TP53, KRAS, NRAS, SMAD4, MET, CTNNB1, PTEN, BRAF, DDR2, EGFR, PIK3CA, NOTCH1, FBXW7, FGFR3, and ERBB2: these have been earlier reported in different tumor tissues. Additionally, 106 novel mutations not reported previously were also detected. One healthy non-smoker subject had a KRAS G12D mutation in EBC DNA.

Our results demonstrate that DNA from EBC of healthy subjects can reveal mutations that could represent very early neoplastic changes or alternatively a normal process of apoptosis eliminating damaged cells with mutations or altered genetic material. Further assessment is needed to determine if NGS analysis of EBC could be a screening method for high risk individuals such as smokers, where it could be applied in the early diagnosis of lung cancer and monitoring treatment efficacy.

No MeSH data available.


KRAS: G12V mutation detected in the EBC sample from a healthy nonsmoking subject
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5392316&req=5

Figure 3: KRAS: G12V mutation detected in the EBC sample from a healthy nonsmoking subject

Mentions: In our study, one specimen exhibited the clinically relevant codon 12 KRAS mutation (G12V) with a mutant allele fraction of 6.8% (Figure 3). This codon mutation was found to be the most frequent mutation in tumor tissue in our previous study of Finnish NSCLC and has also been often described in tissues of lung cancer in other studies [18]. This is in concordance with a recently published study that reported the detection of KRAS G12V mutation in the plasma of three out of six controls, at low concentration (1.25 to 1.87 copies/mL) by using droplet digital PCR [19].


Presence of cancer-associated mutations in exhaled breath condensates of healthy individuals by next generation sequencing
KRAS: G12V mutation detected in the EBC sample from a healthy nonsmoking subject
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5392316&req=5

Figure 3: KRAS: G12V mutation detected in the EBC sample from a healthy nonsmoking subject
Mentions: In our study, one specimen exhibited the clinically relevant codon 12 KRAS mutation (G12V) with a mutant allele fraction of 6.8% (Figure 3). This codon mutation was found to be the most frequent mutation in tumor tissue in our previous study of Finnish NSCLC and has also been often described in tissues of lung cancer in other studies [18]. This is in concordance with a recently published study that reported the detection of KRAS G12V mutation in the plasma of three out of six controls, at low concentration (1.25 to 1.87 copies/mL) by using droplet digital PCR [19].

View Article: PubMed Central - PubMed

ABSTRACT

Exhaled breath condensate (EBC) is a non-invasive source that can be used for studying different genetic alterations occurring in lung tissue. However, the low yield of DNA available from EBC has hampered the more detailed mutation analysis by conventional methods. We applied the more sensitive amplicon-based next generation sequencing (NGS) to identify cancer related mutations in DNA isolated from EBC. In order to apply any method for the purpose of mutation screening in cancer patients, it is important to clarify the incidence of these mutations in healthy individuals. Therefore, we studied mutations in hotspot regions of 22 cancer genes of 20 healthy, mainly non-smoker individuals, using AmpliSeq colon and lung cancer panel and sequenced on Ion PGM.

In 15 individuals, we detected 35 missense mutations in TP53, KRAS, NRAS, SMAD4, MET, CTNNB1, PTEN, BRAF, DDR2, EGFR, PIK3CA, NOTCH1, FBXW7, FGFR3, and ERBB2: these have been earlier reported in different tumor tissues. Additionally, 106 novel mutations not reported previously were also detected. One healthy non-smoker subject had a KRAS G12D mutation in EBC DNA.

Our results demonstrate that DNA from EBC of healthy subjects can reveal mutations that could represent very early neoplastic changes or alternatively a normal process of apoptosis eliminating damaged cells with mutations or altered genetic material. Further assessment is needed to determine if NGS analysis of EBC could be a screening method for high risk individuals such as smokers, where it could be applied in the early diagnosis of lung cancer and monitoring treatment efficacy.

No MeSH data available.