Epidermal growth factor can enhance the developmental competence of pig oocytes matured in vitro under protein-free culture conditions

1999 ◽  
Vol 51 (1) ◽  
pp. 365 ◽  
Author(s):  
L.R Abeydeera ◽  
W.H Wang ◽  
T.C Cantley ◽  
A Rieke ◽  
R.S Prather ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Culture Conditions ◽  
Developmental Competence ◽  
Epidermal Growth

scholarly journals Improvement of developmental competence of aged porcine oocytes by means of the synergistic effect of insulin-like growth factor-1 (IGF-1) and epidermal growth factor (EGF)

2011 ◽  
Vol 50 (No. 7) ◽  
pp. 300-310 ◽  
Author(s):  
I. Petrová ◽  
R. Rajmon ◽  
M. Sedmíková ◽  
Z. Kuthanová ◽  
F. Jílek ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Culture Conditions ◽  
Developmental Competence ◽  
Insulin Like Growth Factor ◽  
Large Numbers ◽  
Complex Process ◽  
Epidermal Growth ◽  
The Many

There is a constant increase in demand for large numbers of high-quality oocytes of domestic mammals for various reproductive biotechnologies. Prolonged in vitro culture represents one of the many possible ways of meeting this demand. However, oocytes further cultured in vitro after reaching the stage of metaphase II undergo the complex process called “ageing”, which decreases the quality of oocytes. The aim of the present study was to improve the culture conditions for in vitro matured porcine oocytes by supplement of the epidermal growth factor (EGF) or the insulin-like growth factor 1 (IGF-1) in order to reduce the adverse effects of ageing. Supplement of either EGF or IGF-1 had no significant effect on the ageing of porcine oocytes. Significant effects were demonstrated when the oocytes were cultured with both EGF and IGF-1. A combination of 10 µg EGF/ml with 25 ng IGF-1/ml was the most effective. Oocytes aged under these conditions retained very good developmental competence. We observed development to the morula (21%) or blastocyst (25%) stage in oocytes aged for 1 day with EGF and IGF-1. Porcine oocytes matured in vitro are more resistant to the ageing when cultured in the presence of both EGF and IGF-1 and these conditions retain an elevated developmental competence for a certain time.  

207 IN VITRO MATURATION TREATMENT AFFECTS DEVELOPMENTAL COMPETENCE OF LAPAROSCOPIC OVUM PICKUP-DERIVED OOCYTES IN FOLLICLESTIMULATING HORMONE-STIMULATED GOATS

2008 ◽  
Vol 20 (1) ◽  
pp. 182 ◽  
Author(s):  
Y. Locatelli ◽  
N. Poulin ◽  
G. Baril ◽  
J.-L. Touzé ◽  
A. Fatet ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Follicular Fluid ◽  
Developmental Competence ◽  
Postmortem Changes ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Epidermal Growth

The aim of the present study was to assess the effect of IVM treatment on the developmental competence of oocytes recovered from repeated laparoscopic ovum pickukp (LOPU) in goats. A total of 94 LOPU sessions were performed on 33 adult goats of the Saanen and Alpine breeds. Females were synchronized (Day 0) during the nonbreeding season by inserting vaginal sponges (45 mg of fluorogestone acetate, Intervet, Boxmeer, The Netherlands). At Day 8, an i.m. injection of 50 μg of cloprostenol (Estrumate; Schering-Plough Animal Health, Pointe-Claire, Quebec, Canada) was administered. Porcine FSH (Stimufol, Merial, Brussels, Belgium, 160 mg/goat) was administered in 5 injections at 12-h intervals, starting on Day 8. The LOPU took place under general anesthesia on Day 11, and follicles ≥2 mm were aspirated with an 18-gauge needle connected to a controlled vacuum system. Vaginal sponges were removed at the time of LOPU. Treatments were repeated 2 times in a 2-week interval scheme (2 goats and 1 goat were excluded from the experiment during the second and third LOPU sessions, respectively). Cumulus–oocyte complexes were washed and evaluated for quality (graded from 1 to 3). Oocytes recovered from unstimulated slaughterhouse-derived ovaries served as a control. Cumulus–oocytes complexes from Grades 1 and 2 were submitted to IVM in TCM-199, supplemented with 100 μm of cysteamine and either 10 ng mL–1 of epidermal growth factor (EGF) or 10% follicular fluid and 100 ng mL–1 of ovine FSH (FF-FSH). Matured oocytes were then submitted to IVF and in vitro development as described by Cognié et al. (2004 Reprod. Fertil. Dev. 16, 437–445). Over the 94 LOPU sessions, 20.4 ± 0.9 follicles were aspirated (mean ± SEM), allowing the recovery of 12.3 ± 0.7 COC per goat and per session, of which 80.1% were suitable for IVM (Grades 1 and 2). Results of in vitro production are detailed in the table. The IVM treatment did not significantly affect cleavage or blastocyst development rates in oocytes derived from slaughterhouse ovaries. Cleavage rates were significantly decreased in LOPU-derived oocytes when compared with control oocytes. For LOPU-derived oocytes, cleavage and final blastocyst development rates were increased significantly and kinetics of embryo development were accelerated when FF-FSH was used during IVM as compared with EGF. The IVM with FF-FSH allowed us to produce 4.1 blatocysts per goat per LOPU session. These results demonstrate the interest in LOPU for goat embryo production once appropriate IVM treatment is used. The difference observed between LOPU and slaughterhouse oocytes in terms of response to IVM treatments may be related to FSH stimulation prior to the LOPU session or to postmortem changes in oocyte responsiveness in the slaughterhouse group. Table 1. Effects of oocyte origin [laparoscopic ovum pickukp (LOPU) or slaughterhouse derived] and maturation treatment [epidermal growth factor (EGF) or follicular fluid (FF)-FSH] on in vitro embryo production (6 replicates)

143 EFFECT OF FSH OR EPIDERMAL-GROWTH-FACTOR-LIKE PEPTIDE SUPPLEMENTATION TO MATURATION MEDIUM ON DEVELOPMENTAL COMPETENCE OF BOVINE OOCYTES DERIVED FROM FULLY DEVELOPED FOLLICLES INDUCED BY SUPER-STIMULATION

2017 ◽  
Vol 29 (1) ◽  
pp. 180
Author(s):  
T. Yamanouchi ◽  
S. Sugimura ◽  
H. Matsuda ◽  
M. Ohtake ◽  
Y. Goto ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Formation Rate ◽  
Calf Serum ◽  
Developmental Competence ◽  
Blastocyst Formation ◽  
Maturation Medium ◽  
Epidermal Growth ◽  
Bovine Oocytes

Bovine oocytes obtained by ovum-pick-up (OPU) following follicle growth treatment (FGT) have improved quality and competence (Imai et al. 2008 Reprod. Fertil. Dev. 20, 182). However, the effect of the presence of FSH or epidermal growth factor (EGF) like peptide during in vitro maturation (IVM) on the developmental competence of FGT oocytes has not been well known. This study was undertaken to examine the developmental competence of FGT oocytes following IVM in the presence of FSH (recombinant human FSH) or EGF-like peptide (amphiregulin; Areg) and IVF. Japanese Black cows (n = 17) were used as donors. Five days after arbitrary OPU (opu group), follicles ≥8 mm in diameter were aspirated again, a controlled internal drug release (CIDR) was inserted into the vagina, and then pFSH was injected twice a day from the evening of Day 6 to the morning of Day 10 with decreasing doses (total of 20 AU; 4, 4, 3, 3, 2, 2, 1, 1 AU/day). On the evening of Day 8, PGF2α (0.5 mg of cloprostenol) was administered. On Day 11, oocytes were aspirated from follicles with ≥5 mm in diameter of the treated donors by OPU (fgt group). The cumulus-oocyte complexes (COC) were cultured in the absence (opu-cont and fgt-cont groups) or presence of 0.1 IU mL−1 FSH (opu-fsh and fgt-fsh groups) or 100 ng mL−1 Areg (opu-areg and fgt-areg groups) in IVM medium (mTCM199 containing 5 mg mL−1 BSA) for 20 to 22 h (1 COC/5 µL, total of 162–171 COC per group), and then co-cultured with 3 × 106 sperm/mL for 6 h. The presumptive zygotes were continued to culture in mCR1aa supplemented with 5% newborn calf serum for 216 h (1 zygote/5 µL) using micro-well culture dishes (Dai-Nippon-Print). When repeating this opu-fgt session in the same cow, an interval at least for 50 days was kept, and the session was performed 28 times. Statistical analysis was carried out by Mann-Whitney’s U-test (between opu and fgt groups) or Steel-Dwass test after Kruskal-Wallis test (among all groups). The number of follicles ≥5 mm increased in the fgt than opu group (17.8 v. 2.9; P < 0.01). The number of COC collected was not different between the opu and fgt groups (23.1 v. 19.6; P > 0.05). The blastocyst formation rate was higher in the fgt than opu group (36.9 v. 23.1%; P < 0.01). Within 6 groups, the blastocyst formation rate was higher in the fgt-fsh (43.3%; P < 0.01) and fgt-areg (39.5%; P < 0.05) groups than the opu-cont (16.3%) group. The rate in the fgt-fsh group was also higher than that in the opu-fsh group (43.3 v. 18.7%; P < 0.01). These results suggested that FGT improved the developmental competence of bovine oocytes, probably through improving the ability of the COC to react against FSH/Areg.

scholarly journals Effect of epidermal growth factor-like peptides on pig cumulus cell expansion, oocyte maturation, and acquisition of developmental competence in vitro: comparison with gonadotropins

Reproduction ◽  
2011 ◽  
Vol 141 (4) ◽  
pp. 425-435 ◽  
Author(s):  
Radek Procházka ◽  
Michal Petlach ◽  
Eva Nagyová ◽  
Lucie Němcová
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Oocyte Maturation ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Human Chorionic Gonadotrophin ◽  
Developmental Competence ◽  
Epidermal Growth

The aim of this work was to assess the FSH-stimulated expression of epidermal growth factor (EGF)-like peptides in cultured cumulus–oocyte complexes (COCs) and to find out the effect of the peptides on cumulus expansion, oocyte maturation, and acquisition of developmental competencein vitro. FSH promptly stimulated expression of amphiregulin (AREG) and epiregulin (EREG), but not betacellulin (BTC) in the cultured COCs. Expression ofAREGandEREGreached maximum at 2 or 4 h after FSH addition respectively. FSH also significantly stimulated expression of expansion-related genes (PTGS2,TNFAIP6, andHAS2) in the COCs at 4 and 8 h of culture, with a significant decrease at 20 h of culture. Both AREG and EREG also increased expression of the expansion-related genes; however, the relative abundance of mRNA for each gene was much lower than in the FSH-stimulated COCs. In contrast to FSH, AREG and EREG neither stimulated expression ofCYP11A1in the COCs nor an increase in progesterone production by cumulus cells. AREG and EREG stimulated maturation of oocytes and expansion of cumulus cells, although the percentage of oocytes that had reached metaphase II was significantly lower when compared to FSH-induced maturation. Nevertheless, significantly more oocytes stimulated with AREG and/or EREG developed to blastocyst stage after parthenogenetic activation when compared to oocytes stimulated with FSH alone or combinations of FSH/LH or pregnant mares serum gonadotrophin/human chorionic gonadotrophin. We conclude that EGF-like peptides do not mimic all effects of FSH on the cultured COCs; nevertheless, they yield oocytes with superior developmental competence.

328 EPIDERMAL GROWTH FACTOR ENHANCES MEIOTIC MATURATION OF CANINE OOCYTES IN THE PRESENCE OF BSA

2006 ◽  
Vol 18 (2) ◽  
pp. 271
Author(s):  
Y.-X. Jin ◽  
X.-H. Shen ◽  
X.-S. Cui ◽  
I.-K. Kong ◽  
N.-H. Kim
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Estrous Cycle ◽  
Oocyte Quality ◽  
Culture Conditions ◽  
Meiotic Maturation ◽  
Lower Percentage ◽  
North Carolina State University ◽  
Epidermal Growth

Despite many attempts to improve the in vitro maturation (IVM) of canine oocytes using various culture conditions, the efficiency of canine IVM remains very low compared with that of other domestic animals. In the present study we examined the effect of (1) ovarian estrous stage on oocyte quality, and (2) epidermal growth factor (EGF) in the presence and absence of macromolecules on the IVM of canine oocytes. Reproductive tracts from normal, client-owned bitches aged approximately 7 months were collected after routine ovariohysterectomy at private veterinary clinics. The ovaries were removed, washed free of blood with PBS, and repeatedly cut with a scalpel blade at room temperature in HEPES-buffered Tyrodes medium containing 0.1% (w/v) polyvinyl alcohol. Each group of 10 cumulus-oocyte complexes (COCs) was matured in North Carolina State University (NCSU)-17 medium supplemented with 1 �g/mL FSH, 10 IU/mL hCG, and 1 �g/mL �-estradiol under paraffin oil for 48 or 72 h at 37�C. Follicular ovaries contained the largest number of oocytes (more than 300), and the number of oocytes e100 �m in diameter was also significantly greater in follicular ovaries than in ovaries at other stages (P < 0.001). Luteal ovaries had the smallest number of both total and large oocytes. After 72 h of culture, significantly more oocytes recovered from follicular ovaries were in GVBD than oocytes from anestrous or luteal ovaries. After 72 h of culture, oocytes recovered from follicular ovaries had a significantly lower percentage reaching the GV stage (59.7 � 5.2%) and more in GVBD (40.3 � 3.1%) compared with both anestrous (GV, 75.1 � 5.2%; GVBD, 24.9 � 3.1%) and luteal (GV, 79.8 � 5.2%; GVBD, 22.0 � 3.1%) ovaries (P < 0.05). However, maturation to MI/MII was not statistically different between the stages of the estrous cycle. BSA or FBS supplementation did not improve meiotic maturation when compared to PVA supplementation. In the presence of 100 nM epidermal growth factor (EGF), either FBS or BSA enhanced the proportion of oocytes in GVBD/MI (59.7 � 2.2% [FBS] and 51.7 � 2.2% [BSA] vs. 36.1 � 2.2% [PVA], P < 0.05). In addition, 0.4% BSA significantly increased the percentage of oocytes in MII (18.5 � 0.8%) compared to 10% FBS (7.1 � 0.8%; P < 0.05). These results suggest that the estrous cycle of bitches influences the meiotic maturation of oocytes cultured in vitro, and EGF increases the meiotic maturation of canine oocytes in the presence of BSA in vitro. This work was funded by a grant from the Ministry of Science and Technology of Korea and the National Research Laboratory Program.

Combination effects of epidermal growth factor and glial cell line-derived neurotrophic factor on the in vitro developmental potential of porcine oocytes

Zygote ◽  
2015 ◽  
Vol 24 (3) ◽  
pp. 465-476 ◽  
Author(s):  
Mehdi Vafaye Valleh ◽  
Mikkel Aabech Rasmussen ◽  
Poul Hyttel
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cell Line ◽  
Oocyte Maturation ◽  
Neurotrophic Factor ◽  
Developmental Competence ◽  
Developmental Potential ◽  
Oocyte Developmental Competence ◽  
Epidermal Growth

SummaryThe developmental potential of in vitro matured porcine oocytes is still lower than that of oocytes matured and fertilized in vivo. Major problems that account for the lower efficiency of in vitro production include the improper nuclear and cytoplasmic maturation of oocytes. With the aim of improving this issue, the single and combined effects of epidermal growth factor (EGF) and glial cell line-derived neurotrophic factor (GDNF) on oocyte developmental competence were investigated. Porcine cumulus–oocyte cell complexes (COCs) were matured in serum-free medium supplemented with EGF (0, 10 or 50 ng/ml) and/or GDNF (0, 10 or 50 ng/ml) for 44 h, and subsequently subjected to fertilization and cultured for 7 days in vitro. The in vitro-formed blastocysts derived from selected growth factor groups (i.e. EGF = 50 ng/ml; GDNF = 50 ng/ml; EGF = 50 ng/ml + GDNF = 50 ng/ml) were also used for mRNA expression analysis, or were subjected to Hoechst staining. The results showed that the addition of EGF and/or GDNF during oocyte maturation dose dependently enhanced oocyte developmental competence. Compared with the embryos obtained from control or single growth factor-treated oocytes, treatment with the combination of EGF and GDNF was shown to significantly improve oocyte competence in terms of blastocyst formation, blastocyst cell number and blastocyst hatching rate (P < 0.05), and also simultaneously induced the expression of BCL-xL and TERT and suppressed the expression of caspase-3 in resulting blastocysts (P < 0.05). These results suggest that both GDNF and EGF may play an important role in the regulation of porcine in vitro oocyte maturation and the combination of these growth factors could promote oocyte competency and blastocyst quality.

267 EFFECT OF FOLLICULAR FLUID CONCENTRATION ON IN VITRO DEVELOPMENT OF PORCINE OOCYTES AND THE EXPRESSION OF GENES RELATED TO CUMULUS EXPANSION AND EMBRYO QUALITY

2013 ◽  
Vol 25 (1) ◽  
pp. 281
Author(s):  
J. E. Park ◽  
H. J. Oh ◽  
M. J. Kim ◽  
G. A. Kim ◽  
E. J. Park ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Polyvinyl Alcohol ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Apoptotic Gene ◽  
Significant Difference ◽  
Epidermal Growth

It is well known that the presence of porcine follicular fluid (PFF) in in vitro maturation media enhances the developmental competence of porcine oocytes. However, it is also suggested that the action of PFF can be modulated positively or negatively by its components. In this study, we investigated the effects of PFF concentration (10 v. 1%) and protein-free media (PFF 0%) on the maturation of porcine oocytes in vitro, and analysed the difference in gene expression in the resulting cumulus cells and blastocysts after parthenogenetic activation. Three groups were tested: 1) 10% PFF: TCM-199 + 10% PFF (n = 638); 2) 1% PFF: TCM-199 + 3.05 mM d-glucose + 1% PFF (n = 418); and 3) 0.1% polyvinyl alcohol: TCM-199 + 3.05 mM d-glucose + 0.1% polyvinyl alcohol (n = 693). Cumulus–oocyte complexes were cultured for 20 to 22 h in the respective media that contained gonadotrophin (1 µg mL–1), epidermal growth factor (10 ng mL–1), cysteine (0.57 mM), sodium pyruvate (0.91 mM), insulin (5 µg mL–1), and 9-cis retinoic acid (5 nM). They were then cultured for an additional 20 to 22 h without hormonal supplements. Data was analysed by one-way ANOVA using the SAS program (SAS Institute Inc., Cary, NC, USA). No significant difference in oocyte maturation rate was observed. However, significantly higher (P < 0.05) proportions of embryos developed in the blastocyst stage when the oocytes were matured in 10% PFF group (45%) than in the 1% PFF group (31.1%). The total cell numbers were not significantly different among groups (52 ± 1.3 v. 54.6 ± 3.1 v. 54.4 ± 2.5, respectively). In addition, the expression of matrix molecule (HAS2, GREM1), steroidogenesis (HSD3B), epidermal growth factor signalling (AREG, BTC), and cell cycle regulator (CCND2) genes were upregulated in the cumulus that was obtained from oocytes that matured in 10% PFF. The expression of the anti-apoptotic gene (BclxL) was upregulated, and the expression of the pro-apoptotic gene (Bax) and metabolism-related genes (GLUT1, LDHA) were downregulated in blastocysts that developed from the 10% PFF group. Therefore, it can be concluded that supplementation of 10% PFF during in vitro maturation improves embryo development by increasing matrix molecules and maturation-enabling factors in the cumulus and by reducing apoptosis. This study was supported by IPET (No. 311011-05-1-SB010), MKE (No. 10033839-2012-21), the Research Institute for Veterinary Science, the BK21 Program, and the TS Corporation.

262 ISOBUTYLMETHYLXANTHINE REVERSIBLY SUPPRESSES SPONTANEOUS AND EQUINE CHORIONIC GONADOTROPIN/EPIDERMAL GROWTH FACTOR-STIMULATED MEIOSIS IN FELINE OOCYTES

2011 ◽  
Vol 23 (1) ◽  
pp. 229
Author(s):  
J. R. Herrick
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Adenylate Cyclase ◽  
Oocyte Maturation ◽  
Mixed Model ◽  
Phosphodiesterase Inhibitor ◽  
Developmental Competence ◽  
Short Term Exposure ◽  
Epidermal Growth

Recent studies have shown that short-term exposure to phosphodiesterase inhibitors (decreased cAMP degradation) or adenylate cyclase stimulators (increased cAMP synthesis) inhibits spontaneous oocyte maturation and improves the developmental competence of oocytes after gonadotropin-induced maturation. A similar approach may improve the developmental competence of in vitro matured feline oocytes. The objectives of this study were to 1) determine if a nonspecific phosphodiesterase inhibitor [isobutylmethylxanthine (IBMX); 100 μM] or an adenylate cyclase stimulator (forskolin; 100 μM) would suppress spontaneous or eCG (1 IU mL–1)/epidermal growth factor (EGF; 25 ng mL–1)-induced maturation of feline oocytes in vitro; 2) evaluate the reversibility of chemically induced meiotic arrest; and 3) assess the developmental competence of meiotically arrested oocytes. Cumulus–oocyte complexes were cultured in modified feline optimized culture medium [6 mM glucose, 0.1 mM cysteamine, 0.6 mM cysteine, 0.5× minimal essential medium (MEM) essential amino acids, 1× MEM vitamins, and 1× insulin-transferrin-selenium; Herrick et al. 2010 Biol. Reprod. 82, 552–562] supplemented with (+) or without (–) IBMX, forskolin, or eCG/EGF (eE). The IBMX decreased (P < 0.05; mixed model ANOVA) the incidence of spontaneous maturation (–eE) after 24 h of culture (84.6%, germinal vesicle, GV; 6.7% metaphase II, MII) compared with control (no supplements) oocytes (18.8% GV, 42.0% MII). Forskolin (–eE) stimulated (P < 0.05) meiosis (6.1% GV, 81.7% MII), so it was not tested in subsequent experiments. The IBMX also inhibited (P < 0.05) eE-induced meiosis compared with control (–IBMX +eE) oocytes after 18 h (87.6 v. 27.1% GV, 2.5 v. 11.0% MII), 24 h (68.0 v. 11.9% GV, 5.6 v. 66.1% MII), and 30 h (58.1 v. 9.4% GV, 14.6 v. 64.4% MII) of culture. To evaluate the reversibility of IBMX, oocytes were cultured +IBMX –eE for 12 h and then transferred to medium –IBMX +eE. After 12 h of culture –IBMX +eE (24 h total), fewer (P < 0.05) IBMX-exposed oocytes were MII (6.6%) compared with control oocytes cultured for 24 h +eE –IBMX (70.2%). The proportion of IBMX-exposed oocytes reaching MII increased (P < 0.05) following 18 h of culture –IBMX +eE (30 h total; 49.4% MII), and by 24 h of culture –IBMX +eE (36 h total; 66.1% MII) was similar (P > 0.05) to the proportion of MII oocytes (78.3%) that had been cultured continuously for 36 h –IBMX +eE. Finally, oocytes were cultured for 0 h or 12 h +IBMX –eE followed by 24 h of culture –IBMX +eE, coincubated with frozen–thawed spermatozoa (5 × 105 sperm mL–1, 22 h), and the resulting embryos cultured (6% CO2, 5% O2) until day 7 post-insemination (Herrick et al. 2007 Biol. Reprod. 76, 858–870). The proportion of oocytes cleaving (83.0 v. 79.9%) and the proportions of oocytes (20.8 v. 18.7%) or embryos (25.2 v. 23.6%) developing to the blastocyst stage were not affected (P > 0.05) by 12 h of IBMX exposure during oocyte maturation. These results demonstrate that IBMX reversibly inhibits both spontaneous and eCG/EGF-induced meiosis in feline oocytes without compromising the oocyte’s developmental competence. This work was supported by the College of Veterinary Medicine’s Companion Animal Memorial Fund at the University of Illinois.

Computational Evaluation and In Vitro Validation of New Epidermal Growth Factor Receptor Inhibitors

2020 ◽  
Vol 20 (18) ◽  
pp. 1628-1639
Author(s):  
Sergi Gómez-Ganau ◽  
Josefa Castillo ◽  
Andrés Cervantes ◽  
Jesus Vicente de Julián-Ortiz ◽  
Rafael Gozalbes
Keyword(s):  
Cell Proliferation ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Binding Affinity ◽  
Cell Lines ◽  
Growth Factor Receptor ◽  
Significant Activity ◽  
Epidermal Growth

Background: The Epidermal Growth Factor Receptor (EGFR) is a transmembrane protein that acts as a receptor of extracellular protein ligands of the epidermal growth factor (EGF/ErbB) family. It has been shown that EGFR is overexpressed by many tumours and correlates with poor prognosis. Therefore, EGFR can be considered as a very interesting therapeutic target for the treatment of a large variety of cancers such as lung, ovarian, endometrial, gastric, bladder and breast cancers, cervical adenocarcinoma, malignant melanoma and glioblastoma. Methods: We have followed a structure-based virtual screening (SBVS) procedure with a library composed of several commercial collections of chemicals (615,462 compounds in total) and the 3D structure of EGFR obtained from the Protein Data Bank (PDB code: 1M17). The docking results from this campaign were then ranked according to the theoretical binding affinity of these molecules to EGFR, and compared with the binding affinity of erlotinib, a well-known EGFR inhibitor. A total of 23 top-rated commercial compounds displaying potential binding affinities similar or even better than erlotinib were selected for experimental evaluation. In vitro assays in different cell lines were performed. A preliminary test was carried out with a simple and standard quick cell proliferation assay kit, and six compounds showed significant activity when compared to positive control. Then, viability and cell proliferation of these compounds were further tested using a protocol based on propidium iodide (PI) and flow cytometry in HCT116, Caco-2 and H358 cell lines. Results: The whole six compounds displayed good effects when compared with erlotinib at 30 μM. When reducing the concentration to 10μM, the activity of the 6 compounds depends on the cell line used: the six compounds showed inhibitory activity with HCT116, two compounds showed inhibition with Caco-2, and three compounds showed inhibitory effects with H358. At 2 μM, one compound showed inhibiting effects close to those from erlotinib. Conclusion: Therefore, these compounds could be considered as potential primary hits, acting as promising starting points to expand the therapeutic options against a wide range of cancers.

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