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Liquid Biopsies

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Introduction

Liquid biopsy is a non-invasive method for detecting intact cancer cells or material released

into body fluids (such as blood and urine) by tumors and/or metastatic disease. It is also known

as fluid phase biopsy or fluid biopsy, and it entails the sampling and examination of non-solid

biological material, primarily blood. Liquid biopsy markers (LBMs) are cancer biomarkers that

include circulating tumor nucleic acids (ctDNA and ctRNA), circulating tumor cells (CTCs),

tumor-derived extracellular vesicles (tdEVs), and tumor-educated platelets (Cowling &

Loshak,2019). It's gaining popularity as a non-invasive alternative to traditional tissue biopsies

for detecting and monitoring cancer biomarkers. The circulating tumor (ctDNA) and cell-free

(cfDNA) subpopulation of ctDNA are detected in exosomes in the blood produced by tumor

cells. Traditional biopsy procedures are mainly utilized for cancer-like disease diagnosis and

monitoring, which is non-invasive. As a result, it allows for a frequency to be used to track

mutations and tumors over time. It's also used to check for effective cancer treatments.

Liquid biopsy is a revolutionary cancer diagnosis and prognosis prediction technology

that uses biofluids like blood, saliva, and urine to detect cancer cells with minimal invasiveness.

Extracellular vesicles (EVs) transfer the molecular payload from donor cells to receiver cells and

are necessary for intercellular communication

Importance of liquid biopsies in the diagnosis of cancer

Fluid biopsy techniques identify the elements in a sample from malignant cells that release

biomarkers and other information such as cell fragments and dead cells or tumor cell necrosis.

Prostate-specific antigen (PSA) prostate screening is one of the most well-known circulating
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protein indicators for cancer diagnosis. Free DNA is utilized to examine liquid biopsies. Cell-free

DNA (cfDNA) is released from tumors and inflammatory tissues during necrosis or apoptosis.

Tumor-derived DNA isolated from plasma can be used to detect mutations in the BRAF proto-

oncogene, KRAS and serine/threonine kinase V600E ( De Rubis et al.,2019). Using circulating

tumor cells, tumor DNA, and other protein markers, a liquid biopsy can help detect and monitor

actionable RASSF1A mutations in lung cancer patients. In several cancer types, including lung

cancer, mutations in EGFR, PIK3CA, BRAF, KRAS, HER2, ALK, PDGFR, and KIT may be

found in circulating tumor DNA. Furthermore, circulating tumor DNA can be exploited with high

specificity and sensitivity. Early on, liquid biopsies based on ctDNA are employed.

Liquid biopsy for mutation detection and monitoring aids in delivering vital information

about a diagnosis, sensitivity, and linked disorders. Tumor mutations must be assessed and

monitored during lung cancer treatment. Patients with specific mutations have a terrible prognosis

and suffer from severe sickness. Precisely identify mutations that necessitate particular treatment

options.

Liquid biopsies require a small amount of blood, urine, or stool. Patients with specific

mutations have a terrible prognosis and suffer from severe sickness. Precisely identify mutations

that necessitate particular treatment options. Liquid biopsies require a small amount of blood,

urine, or stool. Liquid biopsies are less intrusive and provide a reduced procedural risk to the

patient than surgical biopsies, resulting in a potentially less expensive sample collection. Liquid

biopsies can also be done regularly to assess treatment success and/or tumor progression, allowing

for better therapeutic decision-making. As a result, liquid biopsy technology can provide a more

complete picture of disease while also overcoming the spatial limitations of a tissue sample taken

from a single lesion within a single anatomic site ( De Rubis et al., 2019). Cell-free DNA is
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hypothesized to be released from a cancer cell during apoptosis (programmed cell death) or

necrosis (cell death). It can be found in blood and other body fluids such as urine, cerebrospinal

fluid, pleural fluids surrounding the lungs, and saliva, or it can be found in extracellular vesicles

called exosomes. To determine the cell origin's genome or epigenome, 14 cfDNA is examined for

genetic and epigenetic DNA alterations.

Importance of liquid biopsies in the diagnosis of Tuberculosis

Liquid biopsy uses cell-free DNA from the blood. cfDNA opens up new possibilities for

identifying and treating infectious diseases like Tuberculosis. They've become an important

biomarker for quick and non-invasion detection in prenatal, transplantation, and oncology, and

they have the potential to reliably detect a wide range of infectious illnesses, including

Tuberculosis. Using cell-free DNA (cfDNA) in liquid biopsy to detect and treat clinical

infectious diseases brings up new opportunities(Han et al., 2020 ). The successful application of

the cfDNA sequencing test results in accurate non-invasive microbiologic confirmation of this

fastidious organism in less than a month, compared to traditional AFB culture. By detecting

cfDNA in blood and urine, Mycobacterium tuberculosis cfDNA (PCR-based techniques) assists

in determining the presence of infection. Mycobacterium tuberculosis cfDNA assays (PCR-based

procedures) are used to identify cfDNA in blood and urine specimens to diagnose illness. The

results of the mcfDNA sequencing test are used to examine patients with tuberculosis infection.

A plasma mcfDNA sequencing assay successfully delivered accurate non-invasive microbiologic

confirmation of this fastidious pathogen more than one month faster than traditional AFB culture

in invasive Mycobacterium chimera infection cases (Han et al., 2020). Even if the patient had

undergone antibiotic pretreatment, a pathogen cfDNA signal might be identified in plasma.

Mycobacterium tuberculosis infections 54 and aneurysms infected with Mycobacterium Bovis

due to Bacille Calmette-Guérin (BCG) installation show that liquid biopsies are a less invasive

diagnostic and monitoring tool for Tuberculosis. Liquid biopsies are an appealing biomarker for

TB detection,

Comparison to tissue biopsy

Liquid biopsies are increasingly being used to determine what genetic changes or

mutations a tumor has, to inform treatment selection for patients (personalized oncology), to

monitor whether treatments are working and if the minimal residual disease is present after

treatment, and to monitor if cancer has spread to other parts of the body. Liquid biopsies have the

potential to become a cornerstone in oncology, offering some benefits in a range of scenarios.

Two areas with potential benefits include cancer screening and early detection. Liquid biopsies

may detect genetic abnormalities early, allowing for better therapy and outcomes. In the

diagnosis and treatment of advanced cancer, liquid biopsy has shown to be a suitable reference

value. (Kirchner & Jung) (2018) Compared to tissue-based testing, a liquid biopsy revealed

tumor genetic changes with a sensitivity of 0.7, specificity of 0.69, a positive predictive value of

0.86, and a negative predictive value of 0.46.

Liquid biopsy is a simple, quick, non-invasive, and repeatable sample procedure that can

dynamically reflect changes in tumor gene expression profiles and serve as a solid foundation for

customized cancer therapy and early detection. The current accepted diagnostic biomarkers for

screening cancer patients are circulating tumor DNA (ctDNA) and circulating tumor cells

(CTCs). Additional components of liquid biopsies with diagnostic potential include tumor-

derived extracellular vesicles (tdEVs), circulating tumor-derived proteins, circulating tumor

RNA (ctRNA), and tumor-bearing platelets (TEPs).

Ethical issues of liquid biopsies

One of the significant issues in liquid biopsies is the lack of consistency in techniques.

Every technique stage has inconsistencies, from the material used to extract DNA (plasma vs.

serum) to the tools to quantify tumor-associated genetic mutations (digital PCR, NGS,

BEAMing). Multiple techniques are used to preserve and collect samples (blood, plasma, serum,

or DNA) and extract DNA(Jung, 2019). Otherwise, there is a danger of false-negative or false-

positive liquid biopsies. Complex biological data must be created and verified in studies that can

produce therapeutically meaningful outcomes to trigger appropriate treatment decisions and a

deeper understanding of cancer biology and biomarkers. CTCs and ctDNA are often unsuitable

for analysis; therefore, using liquid biopsies for cancer screening and early detection is difficult,

especially in early-stage patients.

Conclusion

Because a patient's fluids are widely available, liquid biopsies are a less stringent

technique of examining the body for various disorders than traditional biopsy methods, allowing

for more general use. It's an exciting new field of study promising better early cancer detection,

treatment modification, and disease recurrence monitoring.

Before referring a patient to an oncologist, liquid biopsies provide a new dimension to the

primary care physician's role in cancer screening and diagnosis. The minimally intrusive nature

of ctDNA profiling enables the diagnosis of malignancy at a microscopic level before radiologic

detection, saving time, money, and risk. Cancers and infectious diseases usually diagnosed late,

such as lung cancer and Tuberculosis, could be detected with a ctDNA assay. Tissue biopsies

reveal a more limited image of each specific malignancy than previously assumed, and
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concurrent liquid biopsies may offer our patients beneficial, lower-morbidity therapy options.

Liquid biopsies for ctDNA have numerous uses during cancer treatment, including dynamic

monitoring of therapeutic response, early resistance detection, and awareness of tumor

recurrence months before clinical relapse. This might return cancer surveillance to the general

internist's practice until a patient's unique tumor marker resurfaces or evolves through repeated

testing.
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References

Cowling, T., & Loshak, H. (2019). An overview of liquid biopsy for screening and early

detection of cancer. CADTH Issues in Emerging Health Technologies.

De Rubis, G., Krishnan, S. R., & Bebawy, M. (2019). Liquid biopsies in cancer diagnosis,

monitoring, and prognosis. Trends in pharmacological sciences, 40(3), 172-186

Han, D., Li, R., Shi, J., Tan, P., Zhang, R., & Li, J. (2020). Liquid biopsy for infectious diseases:

a focus on microbial cell-free DNA sequencing. Theranostics, 10(12), 5501.

Jung, A., & Kirchner, T. (2018). Liquid biopsy in tumor genetic diagnosis. Deutsches Ärzteblatt

International, 115(10), 169.

Martins, I., Ribeiro, I. P., Jorge, J., Gonçalves, A. C., Sarmento-Ribeiro, A. B., Melo, J. B., &

Carreira, I. M. (2021). Liquid biopsies: applications for cancer diagnosis and monitoring.

Genes, 12(3), 349.