Comparison of different tissues on DNA methylation in humans

Changes in DNA methylation have been causally linked with cancer

and provide promising biomarkers for detection in biological fluids
such as blood, urine, and saliva. A number of independent studies
confirm that genome-wide DNA methylation profiles of saliva are more
than 90% comparable to blood, both in adults and in adolescents.
However, based on the analysis of global DNA methylation patterns,
scientists could not completely confirm this. In some studies,
methylation percentages were found significantly lower in saliva
samples compared to blood samples.

But in the field of psychiatric disorders, researchers compared saliva

and blood methylomes with methylation patterns in different brain
tissues. They observed the salivary methylome to be more similar to
methylation patterns in each of the brain regions than methylation in
blood and suggested that DNA methylation profiling using saliva may
offer distinct opportunities for epidemiological and longitudinal studies
of psychiatric traits.

Furthermore, salivary DNA has been used to identify differentially

methylated genes in relation to systemic conditions, such as diabetes
that previously could be detected early with noticing epigenetic
changes in Islets of Langerhans.

Facioscapulohumeral muscular dystrophy (FSHD) is the most

prevalent myopathy that indiscriminately affects males and females of
all ages. It was found that the DNA methylation profile of saliva
reflects FSHD status and it is similar for either cultured cells, tissue,
blood, or saliva samples.
In addition, salivary analysis has been shown to be a useful
diagnostic tool in the field of cancer research cancer‐associated
genes in the saliva of patients with oral cancer and reported that
methylation array analysis of saliva can produce a set of cancer‐
related genes that can be used as a composite biomarker for the early
detection of oral cancer.

Moreover, gene promoter methylation analysis of a test panel in

salivary DNA was able to detect the early stages of head and neck
squamous cell carcinoma that is one of the very prominent types of
skin cancer.

Salivary DNA methylation analyses have also been applied for distant
malignancies such as breast cancer. Several salivary DNA
methylation markers have been identified in breast cancer‐related
genes and were associated with risk factors for breast cancer
development. Overall, these data indicate that saliva is a useful
source of DNA for detecting differential methylation marks in a non‐
invasive manner in vulnerable groups.

DNAm tends to be highly tissue- or cell-type specific. methylation

patterns from airway epithelial cells (AECs), nasal epithelial cells
(NECs) might serve as better biomarkers for the diagnosis of asthma
compared with methylation patterns derived from peripheral blood
mononuclear cells (PBMCs).

In some studies it was found that methylation patterns from AECs and
NECs might serve as better biomarkers for the diagnosis of asthma
compared with methylation patterns derived from PBMCs.

Recently, in a study, a case–control design was used to analyze and

compare DNA methylation patterns in blood and saliva in individuals
with respiratory allergies. When comparing respiratory allergy cases
with healthy controls, 485 and 437 differentially methylated sites were
identified in saliva and PBMC, respectively, of which 216 were in
common and showed the same polarity in blood and saliva.

For urological cancers, urine is in many situations the preferred “liquid

biopsy” source because it contains exfoliated tumor cells and cell-free
tumor DNA and can be obtained easily, noninvasively, and
repeatedly.

Urine-based DNA tests for urological cancer can be divided into two
categories depending on the a priori availability of information on the
patient’s tumor DNA. For detection of recurrence and evaluation of
treatment response, DNA from the original tumor can be analyzed to
identify specific alterations that may serve as “personalized”
biomarkers. For other applications, such as initial examination of
patients with symptoms of urological cancer, the genetic and
epigenetic makeup of the possible tumor is unknown. In these
situations, there is a need for a “universal” or “generic” test that can
detect, in principle, any cancer. Because no genetic or epigenetic
alteration is present in all cases of a urological cancer type, it is
necessary to use a combination of biomarkers. Urine is tested for
early detection of a number of cancers urological cancer, bladder
cancer, prostate cancer, upper urinary tract cancer, renal cancer.

Another factor that should be considered when designing urine-based

assays for urological cancer is that the concentrations of cells and
DNA in urine are not constant. Shedding of cells and the release of
DNA through apoptosis or necrosis are stochastic and depend on
several factors, including the size and location of the tumor. Studies
over more than a decade have demonstrated the great potential of
DNA methylation biomarkers for urine-based detection of urological
cancer. However, the bewildering number of biomarkers currently
under evaluation and the great variability in biomarker performance
across studies hamper successful translation into clinically useful
tests.

Some paper findings suggest that promoter hypermethylation in urine

or serum can be detected in the majority of renal cancer patients. This
noninvasive high-throughput approach needs to be evaluated in large
studies to assess its value in the early detection and surveillance of
renal cancer.

Abnormal promoter methylation in saliva DNA was found in all tumor

stages and more frequently in tumors located in the oral cavity.
Moreover, none of the saliva from patients with methylation-negative
tumors displayed methylation of any marker. In one recent study, of
30 saliva samples from healthy control subjects (15 smokers and 15
nonsmokers), only one sample from a smoking patient was positive
for DNA methylation at two target genes. Detection of aberrant
promoter hypermethylation patterns of cancer-related genes in the
saliva of head and neck cancer patients was found feasible.

Our results are in general agreement with the detection of promoter

hypermethylation in the serum DNA of head and neck squamous cell
carcinoma and in serum and bronchoalveolar fluid of lung cancer
patients. Recent publications have demonstrated the presence of
promoter hypermethylation in the serum DNA of lung, liver, breast,
and head and neck cancer patients. In addition, genetic changes
identified in tumors can also be detected in bodily fluids in direct
contact with the neoplastic site, including the urine of bladder cancer
patients
Lung cancer has a high incidence rate. Mutations identified in the
EGF receptor (EGFR) are the tumor-specific biomarkers for non-small
cell lung carcinoma (NSCLC). A novel core technology known as
electric field-induced release and measurement relies on a
multiplexible electrochemical sensor that can detect EGFR mutations
in bodily fluids was shown to be effective, accurate, rapid, and cost-
effective for the detection of EGFR mutations in the saliva of patients
with NSCLC.

C-reactive protein (CRP) was the most predictive biomarker of acute

myocardial infarction. Acute myocardial infarction was predicted by a
combination of electrocardiogram and CRP levels with 80.0%
sensitivity and 100% specificity. These data demonstrated the
potential use of salivary biomarkers with electrocardiogram for the
diagnosis of acute myocardial infarction. Moreover, the levels of α-2-
HS-glycoprotein in saliva decreased in patients with CVD, which
indicates that the peptidome might provide a potential way for the
early diagnosis of patients with CVD.

In rodent models of pancreatic cancer, vesicles similar to exosomes

can carry and transport tumor-specific biomarkers into the saliva.

MiR-141 and miR-21 are two tumor biomarkers; the former is

significantly elevated in patients with advanced-stage prostate cancer,
whereas the latter is overexpressed in early-stage prostate cancer. It
has been demonstrated that the expression of miR-21 and miR-141 in
the saliva can be detected by nano-graphene oxide. This is expected
to be a non- or minimally invasive approach to diagnose early-stage
prostate cancer.

In the saliva of patients with gastric ulcers and chronic gastritis,

Helicobacter pylori DNA can be detected to identify H. pylori
infection.50 Significant correlations were found between salivary
caffeine clearance and liver diseases. Thus, saliva can be used as an
effective biochemical parameter for the diagnosis of chronic liver
diseases (CLDs) and assessment of residual liver function in CLD.

Helicobacter pylori (H.pylori), which is known to cause inflammation of

the stomach lining, can also lead to gastric cancer. Two metabolites
of H.pylori can be detected in saliva with good sensitivity. H.pylori,
causing the stomach lining inflammation, can also lead to gastric
cancer. Zilberman et al. detected clinically relevant levels of two
metabolites of H.pylori, NH3 and CO2, in saliva, which provides a
platform for cross-reactive sensitivity and allows detection of salivary
CO2 and NH3 at ppm levels

The saliva of patients with chronic renal failure presented significantly

higher levels of NO.

This study developed a composition for stable storage of saliva RNA

at room temperature and confirmed that saliva can be used as a
sampling source for the detection of leukemic fusion transcripts.
Saliva offers advantages over blood and bone marrow for its non-
invasive nature, easy storage and transportation, cost-effectiveness,
and safe handling.

In particular, two studies have validated the feasibility of detecting

increased levels of miRNAs in the saliva of CRC patients. In one of
these studies, miR-21 levels were found to be increased in both
plasma and saliva of patients with stage II-IV CRC compared to those
of healthy individuals. Importantly, the sensitivity and specificity of
CRC identification were higher when analyzing saliva samples.
Recently, miR-21 expression was assessed in peripheral blood and
saliva samples obtained from patients with CRC at different stages
and healthy controls. Although miR-21 levels in both saliva and
plasma, showed diagnostic value for CRC screening, the analysis of
saliva demonstrated higher sensitivity and specificity than blood.

Altered epigenetic profiles are a feature of intestinal diseases,

including ulcerative colitis and Crohn’s disease. The results from
some studies provide a framework for the use of saliva in DNA
methylation research studies in intestinal conditions. they suggest that
saliva has the potential to be used as an alternative for intestinal
mucosa.

Adenoid cystic carcinoma (ACC) is a rare form of adenocarcinoma, a

type of cancer that begins in glandular tissues. In primary Adenoid
Cystic Carcinoma (ACC) tissues, AQP1 is both hypomethylated and
overexpressed at mRNA and protein levels when compared to normal
salivary tissue

Morning salivary cortisol is as good as serum as a screening test for

patients with Addison's disease and nighttime salivary cortisol is more
adequate than serum in the screening of Cushing's syndrome.
Salivary cortisol reflects the free and biologically active fraction of
cortisol in serum. We evaluated the usefulness of salivary cortisol
measurements in the assessment of Cushing's syndrome and adrenal
insufficiency. Plasma and salivary cortisol, as well as 6beta-
hydroxycortisol determinations, were performed by
radioimmunoassay after extraction with ethyl acetate followed by
chromatographic separation using a modified paper chromatographic
system. Samples were obtained from 36 control subjects and 37
patients with non-hyperfunctioning adrenocortical adenomas in the
morning at 8 a.m. in that study, results suggest that plasma and
salivary 6beta-hydroxycortisol determinations may precisely detect
not only overt increases of cortisol secretion in patients with Cushing's
syndrome but also mild glucocorticoid overproduction presumably
present in patients with non-hyperfunctioning adrenocortical tumors.

So, in conclusion, it can be said one can use the DNA methylation
from saliva and/or blood as a reference for the specific methylation
patterns or biomarkers for most of the diseases of other tissues. And
in some cases, saliva tests are more preferable to blood tests as they
are non-invasive — the test results “may be more accurate in that less
stress on the system during the production of the specimen means
less interference with the factors being tested.” They are easier to
obtain and less painful — no syringes, no scared patients the same
time being less expensive.