Introduction
Extracellular vesicles (EVs) are nanoscale lipid-enclosed vesicles released by all cells. For a long time, the scientific community overlooked the importance of EVs since they were only assumed to participate in the control of cellular homeostasis, i.e., to function as “waste carriers” [1]. However, after discovering EVs ability to transfer active molecules such as RNA and proteins, EVs have gained strong interest as potential key players in different fields such as normal physiology, pathology including cancer, neurodegenerative disorders, and drug delivery strategies to mention a few [[2], [3], [4]]. EVs are broadly classified based on their size and route of biogenesis, such as exosomes which originate from the endolysosomal pathway; microvesicles which bud from the plasma membrane and apoptotic bodies which are released from dying cells [5]. Due to the difficulties in selective isolation, EVs are often classified as small EVs (sEVs; 100–200 nm) and medium/large EVs (m/lEVs; >200 nm) [6]. It is increasingly evident that there exist subclasses of EVs in each group making them phenotypically and functionally more heterogeneous [5,6]. Identifying such subpopulations of EVs is currently one of the major efforts in the EV field which could improve on their biomarker utility [7].
The treatment regimens for non-small cell lung cancer (NSCLC) patients with advanced or metastatic disease have changed during the last decade where molecular profiling has identified oncogenic genomic alterations e.g.epidermal growth factor receptor (EGFR; e.g. E19del, exon 20 T790 M, and exon 21 L858R mutations), Kirsten rat sarcoma virus (KRAS; e.g. G12C mutation) and v-raf murine sarcoma viral oncogene homolog B1 (BRAF) or gene fusions/rearrangements e.g., echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK), allowing for precision cancer treatments using receptor tyrosine kinase (RTK) inhibitors (TKIs) [[8], [9], [10]]. Some NSCLC patients have also benefited from re-activation of the immune system towards the tumor using so-called immune checkpoint inhibitors (ICI) acting on the programmed cell death protein 1/Programmed death-ligand 1 (PD-1/PD-L1) axis [11]. About 12–30% of all NSCLC are driven by mutated EGFR. Such cases are treated with EGFR-TKIs. These include 1st generation gefitinib and erlotinib but lately also with 2nd generations TKIs such as afatinib and dacomitinib, but importantly also by the 3rd generation osimertinib. EGFR-TKIs may allow a long progression-free survival (PFS), with osimertinib treatment showing the longest PFS [9]. However, as responses to EGFR-TKIs are heterogenous among NSCLC patients, there is a need for ways to early monitor changes in treatment response in a non-invasive way e.g., in patient blood samples.
Profiling of EVs for early lung cancer diagnostics using bulk EVs samples for protein or RNA analyses has deciphered certain markers [[12], [13], [14], [15]]. Multiple studies in different tumor types have also demonstrated that EGFR signaling may influence the EVs released in terms of tetraspanins CD9, CD63, and CD81 expression as well as other proteins [16,17]. It has been shown that EVs contain various immune signaling components including programmed death-ligand 1 (PD-L1), which could contribute to an immune suppressive phenotype seen among NSCLC cases [18,19]. Yet, most of these EV studies have focused on bulk analyses of EVs, thereby possibly missing important signatures hidden in the heterogenous EV population [12,18,20,21].
Using a fluorescence-based single-EV approach in this study, we demonstrate that the membrane proteins of EVs reveal interesting differences which, if validated in clinical cohorts, can potentially be used for early detection of treatment response of targeted cancer therapies using liquid biopsy. For this purpose, we analyzed five membrane proteins (CD9, CD81, EGFR, human epidermal growth factor receptor 2 (HER2), and PD-L1) on individual EVs extracted from cell culture media of the EGFR-mutant NSCLC cell line H1975, both prior and post treatment with EGFR-TKIs erlotinib or osimertinib. The treatment selection was motivated to highlight the differences in EV surface protein expression when the tumor is refractory versus responsive to the applied drugs. To further increase our understanding, we compared the results with those obtained in EVs isolated from cell culture media after cisplatin chemotherapy. The results indicate that in contrast to erlotinib and cisplatin, osimertinib treatment increased a subset of the HER2 and PD-L1 positive EV population. However, the average expression of the two proteins on individual EVs was found to drop after osimertinib treatment. Differences across the samples were also observed in the distribution of expression levels on individual EVs positive for either HER2, EGFR, or PD-L1. On the other hand, as a common effect of both the EGFR-TKIs, the EGFR positive EV population was increased after the treatments whereas the average EGFR expression level per EV was decreased.
Section snippets
Reagents
High purity deionized water (DIW) with a resistivity of 18 MΩ cm was used throughout all the experiments. Casein (C5890) in powder, phosphate-buffered saline (PBS, P4417) in tablets, and poly-l-lysine (PLL, P2636) were purchased from Sigma-Aldrich (Burlington, MA, US). Anti-CD9 antibody conjugated with VioBlue (#130-118-809) was bought from Miltenyi Biotec (Bergisch Gladbach, NW, DE), while anti-EGFR R-phycoerythrin (R-PE, #1P-680-T100), anti-Human CD274/PD-L1 APC (#1A-177-T100), and anti-Human
Characterization of H1975 cells and EVs from cell culture media
The cell viability of the H1975 cell line after exposure to the EGFR-TKIs erlotinib, osimertinib or cisplatin chemotherapy is presented in Fig. 2A. Images of cell morphology are presented in supporting information (Figure S2). Morphology of erlotinib-treated cells has already been published [26]. The results agree with our earlier reports that osimertinib at this concentration causes a 50% reduction in cell viability, while erlotinib at 1 μM only had a minor effect on the cells [26]. In
Discussion
Targeted agents for metastatic NSCLC have changed the treatment regimen and patient outcome. Thus, forEGFR-mutant cases, the introduction of EGFR-TKIs has improved the survival rate [30]. Yet responses are often heterogenous, and development of resistance is common even with 3rd generation osimertinib. Osimertinib is currently also being explored as an adjuvant treatment for early NSCLC cases [9,10]. As a result, there is a need for non-invasive diagnostic and treatment monitoring approaches to
Conclusions
In summary, we demonstrated that a single-EV analysis of membrane proteins holds the potential for monitoring EGFR-TKI response in NSCLC patient plasma samples albeit this must be verified in a clinical cohort with differential osimertinib response. Thus, one limitation of our study is that we conducted the experiments with one EGFR-mutant NSCLC cell line, albeit three different treatment types. The chosen cell line was resistant to erlotinib and responsive to osimertinib or cisplatin. Thus,
Credit author statement
Fredrik Stridfeldt: conceptualization, methodology, software, validation, formal analysis, investigation, data curation, writing-original draft, writing-review & editing, visualization; Sara Cavallaro: conceptualization, methodology, writing – review and editing.; Petra Hååg: methodology, validation, investigation, resources, writing; Rolf Lewensohn: funding acquisition, writing; Jan Linnros: funding acquisition, conceptualization, supervision.; Kristina Viktorsson: conceptualization,
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
This project was supported by a grant from the Erling Persson Family Foundation. A.D., S.C., and F.S., acknowledge also the grant funded by the Swedish Research Council (Contract No. 2016–05051 and 2018–06228). The biological part of the study was granted to R.L, K.V and P.H from the Swedish Cancer Society (contract no. CAN 2018/597 and CAN2021/1469), the Stockholm Cancer Society (contract no. 201202, 191293, 221212 and 221383) and Stockholm County Council (contract no. 909121 and 750032). The
© 2023 Published by Elsevier B.V.
FAQs
What is epidermal growth factor receptor cancer? ›
A gene that makes a protein that is involved in cell growth and cell survival. Mutated (changed) forms of the epidermal growth factor receptor gene and protein have been found in some types of cancer, including non-small cell lung cancer. These changes may cause cancer cells to grow and spread in the body.
What is the EGFR test for lung cancer? ›An EGFR mutation test may be ordered by itself or as part of a panel (a series of tests to detect mutations in other genes such as KRAS, ALK and ROS1). Each of these tests may be used to help determine whether a person's lung cancer will respond to targeted therapy and which type will be of more benefit.
What does EGFR positive mean? ›A positive test for the EGFR biomarker means that you are eligible for targeted therapy. Targeted therapy drugs keep cancer from growing and spreading with less harm to cells that aren't cancer. The drugs for EGFR+ cancer are tyrosine kinase inhibitors. They aim to block the EGFR protein.
How do EGFR inhibitors work? ›A substance that blocks the activity of a protein called epidermal growth factor receptor (EGFR). EGFR is found on the surface of some normal cells and is involved in cell growth. It may also be found at high levels on some types of cancer cells, which causes these cells to grow and divide.
What is the role of epidermal growth factor receptor in lung cancer? ›EGFR (epidermal growth factor receptor) is a protein on cells that helps them grow. A mutation in the gene for EGFR can make it grow too much, which can cause cancer.
What is epidermal growth factor receptor in Nsclc? ›Epidermal growth factor receptor (EGFR), is one of several somatic mutations, in NSCLC (6), which is seen more frequently in certain population groups. This population group is classically described as Asian, non-smoking females with adenocarcinoma (7-9).
What is EGFR mutation in non-small cell lung cancer? ›Epidermal growth factor receptor (EGFR) mutations are the most common oncogenic drivers in non-small-cell lung cancer (NSCLC). Significant developments have taken place which highlight the differences in tumor biology that exist between the mutant and wild-type subtypes of NSCLC.
What is the best treatment for EGFR positive lung cancer? ›If you have advanced, or stage IV, NSCLC and it's EGFR-positive, your doctor will likely treat your cancer with a targeted therapy. These medicines are called EGFR inhibitors, or EGFR targeted tyrosine kinase inhibitors (TKIs). These drugs block the EGFR signal that makes your cancer cells grow.
What is the new treatment for EGFR positive lung cancer? ›Way takes Tagrisso, a targeted therapy for non-small cell, early-stage epidermal growth factor receptor (EGFR)-positive lung cancer, where a mutation in the tumor causes cancer to spread quickly.
What happens if eGFR is high? ›An eGFR below 60 for three months or more, or an eGFR above 60 with kidney damage (marked by high levels of albumin in your urine) means chronic kidney disease. Your healthcare team will want to find the cause of your kidney disease and continue to check your kidney function to help plan your treatment.
What cancers have eGFR mutations? ›
Overview. EGFR Mutation is present in 5.48% of AACR GENIE cases, with lung adenocarcinoma, conventional glioblastoma multiforme, glioblastoma, colon adenocarcinoma, and non-small cell lung carcinoma having the greatest prevalence [4].
Should eGFR be high or low? ›A normal eGFR is 60 or more. If your eGFR is less than 60 for three months or more, your kidneys may not be working well.
What is the life expectancy of Stage 4 Tagrisso? ›reTHINK stage 4
The median progression-free survival was 18.9 months for TAGRISSO vs 10.2 months for erlotinib or gefitinib. In the same clinical study, median overall survival was 38.6 months for TAGRISSO vs 31.8 months for erlotinib or gefitinib.
also help to slow or stop your lung cancer from growing or help to shrink the tumour. TAGRISSO has been shown to produce effects on the tumour within 6 to 12 weeks of starting therapy.
What is the survival rate for EGFR patients? ›In the last 10 years, first-generation EGFR-TKIs, such as gefitinib, erlotinib, and icotinib, have become indispensable treatments for EGFR mutation-type advanced NSCLC. The 5-year survival rate of patients with EGFR-mutant metastatic lung adenocarcinoma treated with EGFR-TKIs is 14.6%.
How long can you live with EGFR lung cancer? ›In this study, the OS of EGFR-mutated NSCLC was 36.9 months, which was longer than that in the IPASS study, at 21.6 months [23]. The reason may be that 16% (26/155) of patients took osimertinib. In addition, Jessica et al. reported that the OS in patients with EGFR-mutated lung cancer was 30.9 months.
What causes EGFR mutation in lung cancer? ›An EGFR mutation occurs when there is an error in the DNA that makes up the protein. These errors are also a type of biomarker. A biomarker is a biological molecule that can be an indicator of a certain condition or disease, according to the National Cancer Institute .
What is the significance of epidermal growth factor? ›Epidermal growth factor (EGF) is a 53-amino-acid cytokine which is proteolytically cleaved from a large integral membrane protein precursor2. EGF acts to stimulate growth of epithelial cells, accelerates tooth eruption and eyelid opening in mice, inhibits gastric acid secretion and is involved in wound healing3.
What is anti EGFR epidermal growth factor? ›The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor involved in the proliferation and survival of cancer cells. EGFR is the first molecular target against which monoclonal antibodies (mAb) have been developed for cancer therapy.
What is epidermal growth factor EGF and what function does it perform? ›A protein found on certain types of cells that binds to a substance called epidermal growth factor. The epidermal growth factor receptor protein is involved in cell signaling pathways that control cell division and survival.
What are epidermal growth factor receptor drugs? ›
Epidermal growth factor receptor (EGFR, also known as ErbB-1 or HER-1) inhibitors are medicines that bind to certain parts of the EGFR and slow down or stop cell growth. EGFR is a protein that is found on the surface of some cells that causes cells to divide when epidermal growth factor binds to it.
How curable is non-small cell lung cancer? ›Because stage 0 NSCLC is limited to the lining layer of the airways and has not invaded deeper into the lung tissue or other areas, it is usually curable by surgery alone. No chemotherapy or radiation therapy is needed.
What is the poor prognosis of non-small cell lung cancer? ›The 5-year survival rate for men is 18%. The 5-year survival rate for women is 25%. The 5-year survival rate for NSCLC is 26%, compared to 7% for small cell lung cancer. However, it is important to note that survival rates depend on several factors, including the subtype of lung cancer and the stage of disease.
What stage is non-small cell lung cancer? ›Stage IV non-small cell lung cancer (NSCLC) is the most advanced form of lung cancer. In stage IV, the cancer has metastasized, or spread, to the lining of the lung or other areas of the body.
What is the first-line treatment for EGFR? ›The epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) gefitinib, erlotinib and afatinib are currently recommended for first-line treatment of patients with advanced non-small cell lung cancer (NSCLC) harboring EGFR-TKI-sensitizing mutations (EGFRm) (1,2).
What is the best treatment for lung cancer in one lung that hasn t spread? ›Surgery. NSCLC that hasn't spread and SCLC that's limited to a single tumor can be eligible for surgery. Your surgeon might remove the tumor and a small amount of healthy tissue around it to make sure they don't leave any cancer cells behind.
What is the first-line treatment for EGFR NSCLC? ›These include efficacy, toxicity, quality of life, and cost–benefit analysis. The results of IRESSA Pan-Asia Study and EURopean TArceva vs. Chemotherapy trials established gefitinib/erlotinib as the first-line treatment of EGFR-mutant NSCLC. However, patients invariably progress after 9–12 months.
What is the new treatment for non-small cell lung carcinoma? ›Atezolizumab is approved to treat some people with non-small cell lung cancer after surgery. The HER2 mutations seen in lung cancer make the HER2 protein activated all the time, Dr.
What drugs treat EGFR non-small cell lung cancer? ›EGFR inhibitors used for squamous cell NSCLC
Necitumumab (Portrazza) is a monoclonal antibody (a lab-made version of an immune system protein) that targets EGFR. It can be used with chemotherapy as the first treatment in people with advanced squamous cell NSCLC. This drug is given as an infusion into a vein (IV).
In 2021, the U.S. Food and Drug Administration (FDA) approved targeted therapy drugs for this specific NSCLC mutation for the first time. Targeted therapy is a cancer treatment approach that uses drugs to target specific genes and proteins that are involved in the growth and survival of cancer cells.
Does drinking water increase GFR? ›
Volume of water intake positively correlated to estimated glomerular filtration rate (eGFR), and negatively correlated to urinary albumin to creatinine ratio (UACR), as well as plasma osmolality and urine osmolality, although the correlations were weak.
Can eGFR go back to normal? ›While it's possible to improve your GFR, you're more likely to do so with acute kidney injuries rather than with chronic kidney disease. For most people with chronic disease, positive lifestyle changes may help slow the loss of kidney function.
Can dehydration lower eGFR? ›Dehydration does cause the serum creatinine to rise and the estimated glomerular filtration rate (eGFR) will, accordingly, fall. The degree of change is generally proportional to the degree of dehydration. Severe dehydration can actually cause acute kidney injury and may lead to a need for dialysis therapy.
What is the commonest EGFR mutation? ›L858R are the most common activating mutations in EGFR, accounting for approximately 90% of all EGFR mutations in NSCLC [5, 6]. EGFR tyrosine kinase inhibitors (TKIs) have shown profound clinical benefits and are thus used as the first-line treatment in EGFR-mutated NSCLC patients [7,8,9,10,11,12].
Does Tagrisso shrink tumors? ›77% of those taking TAGRISSO saw their brain tumors shrink. 18% saw their brain tumors completely disappear.
What is the most frequent mutation in EGFR? ›The two most common EGFR mutations are short in-frame deletions of exon 19 and a point mutation (CTG to CGG) in exon 21 at nucleotide 2573, which results in substitution of leucine by arginine at codon 858 (L858R).
What is the best thing to drink for your kidneys? ›Water: Water is simply the best drink you can have! Water is a zero-calorie, perfectly hydrating, cheap drink. If you are in the earlier stages of kidney disease, choosing water most of the time to quench your thirst will keep your body and kidneys functioning well.
What is a dangerously low eGFR? ›Stage 5 (eGFR below 15) is a sign of kidney failure. It means you have less than 15% kidney function. This stage is the most serious and can be life-threatening. You will need dialysis (a machine to filter your blood) or a kidney transplant.
What food should be avoided if creatinine is high? ›Kapoor says a healthy diet can aid in lowering creatinine levels and one must avoid foods like red meat, salty meals, white bread, processed foods, sugary foods, caffeine, canned vegetables, and foods that may contain high quantities of protein if diagnosed with high levels of creatinine in the blood.
Is Tagrisso better than chemo? ›Tagrisso is also an effective second-choice option for people that are resistant to other oral EGFR inhibitors and move on to chemotherapy. In a different study, Tagrisso lengthened the time without seeing cancer growth by about 6 months compared to chemotherapy.
How long can people live on Tagrisso? ›
Tagrisso demonstrated 5.5-year median disease-free survival in the adjuvant treatment of patients with EGFR-mutated lung cancer.
Is Tagrisso approved for early stage NSCLC? ›AstraZeneca's Tagrisso (osimertinib) has been approved in the European Union (EU) for the adjuvant treatment of adult patients with early-stage (IB, II and IIIA) epidermal growth factor receptor-mutated (EGFRm) non-small cell lung cancer (NSCLC) after complete tumour resection with curative intent.
Do you lose hair with Tagrisso? ›For example, at week 2, 13% of patients taking Tagrisso reported Hair Loss (A little bit). The range of patients who had any Hair Loss during the first 24 weeks of treatment with Tagrisso was between 7% - 13%.
Who should not take Tagrisso? ›heart problems, including heart failure.
TAGRISSO may cause heart problems that may lead to death. Your healthcare provider should check your heart function before you start taking TAGRISSO and during treatment as needed.
Caffeine osimertinib
Osimertinib may reduce the blood levels and effects of caffeine. Contact your doctor if your symptoms worsen or your condition changes during treatment with these medications.
In the general population, approximately 38% of adults aged 70 or older have an estimated glomerular filtration rate (eGFR) < 60 ml/min/1.73 m2, with most having moderate reductions in eGFR in the 30-59 ml/min/1.73 m2 range (1).
What is the lowest GFR you can live with? ›85-90% of kidney function is gone. GFR falls below 15. Kidneys don't work well enough to keep you alive.
Is Stage 2 eGFR reversible? ›Treatment for stage 2 kidney disease. Once kidney damage occurs, you can't reverse it. However, you can prevent further progression. This involves a combination of lifestyle changes and medications to help treat the underlying causes of stage 2 CKD.
What does EGFR mean in breast cancer? ›The epidermal growth factor receptor (EGFR) is one of the first identified important targets of these novel antitumor agents. Approximately half of cases of triple-negative breast cancer (TNBC) and inflammatory breast cancer (IBC) overexpress EGFR.
Is epidermal growth factor good or bad? ›"EGF in skin care helps combat the visible signs of aging by strengthening the skin barriers, boosting hydration, minimizing the appearance of fine lines and wrinkles, and helping support the skin's natural collagen production, keeping the skin looking youthful and healthy," says Dr. Örvar.
What does the epidermal growth factor do? ›
A protein made by many cells in the body and by some types of tumors. It causes cells to grow and differentiate (become more specialized). It is a type of growth factor and a type of cytokine.
What do growth factors do in cancer? ›Growth factors can also influence normal cell differentiation, and constitutive activation of growth-promoting pathways in cancer cells can modulate the cell phenotype as well. Paracrine actions of growth factors and cytokines may also influence the stepwise series of genetic events that lead to malignancy.
Is high eGFR good or bad? ›An eGFR higher than 60 means you have at least 60% kidney function. Generally, the higher the number, the better your kidney function.
Does a low eGFR mean cancer? ›Lower eGFR strata were significantly associated with higher risk of skin, urogenital, prostate, and hematologic cancers.
Should I worry if my eGFR is high? ›An eGFR below 60 for three months or more, or an eGFR above 60 with kidney damage (marked by high levels of albumin in your urine) means chronic kidney disease. Your healthcare team will want to find the cause of your kidney disease and continue to check your kidney function to help plan your treatment.
What are the side effects of epidermal growth factor receptor therapy? ›Other cutaneous side effects include xerosis and hair and nail changes. The onset of eruption is usually within one to three weeks after starting therapy, although in some cases it may occur much later. All dermatologic side effects are reversible and generally resolve after adequate therapy.
Are growth factor serums worth the money? ›Are EGF serums worth it? Epidermal Growth Factors (EGF) are derived in a lab from human sources, contributing to their higher cost. EGF products contain countless growth factors, increasing potency and effectiveness. Growth factors derived from human sources are the most effective and are definitely worth the price.
What are the 5 epidermal growth factors? ›- Basic Fibroblast Growth Factor.
- Eicosanoid Receptor.
- Epidermal Growth Factor Receptor.
- Cell Signaling.
- Transforming Growth Factor Beta.
- In Vitro.
- Cell Proliferation.
- Transforming Growth Factor Alpha.
EGFR mutations usually occur during the patient's lifetime, although in a small number of cases the EGFR mutation can be inherited from a parent. EGFR mutations are found more frequently in never-smokers but can also occur in former smokers.
What activates epidermal growth factor? ›According to the “ligand-induced dimerization model”, EGFR is activated by the ligand-induced dimerization of the receptor monomer, which brings intracellular kinase domains into close proximity for trans-autophosphorylation to initiate downstream signaling cascades.
How long does it take EGF to work? ›
How long will it take to see results? Used as directed, the beneficial effects of EGF can be perceivable within days. Improvements in lines and wrinkles are often noticed within 1-2 weeks, however, depending on your skin condition, it may take a couple of months to see more dramatic results.
What are the 3 factors which can increase the chances of cancerous growth? ›Age, weight, exposure to carcinogens, and genetics can increase the risk of developing cancer.
What is the most significant factor for developing cancer? ›Tobacco smoking is overwhelmingly the most significant risk factor for cancer and across the board for chronic diseases. Diet, exercise, and alcohol use also cut across the diseases, and they are significant contributors to cancer, but more significant to other conditions.
What are the 4 growth factors? ›skin cells (epidermal growth factor, EGF) nerve cells (nerve growth factor, NGF) connective tissue or mesenchymal cells (fibroblast growth factor, FGF) thrombus-forming cells that line blood vessels (platelet- derived growth factor, PDGF)