Gulnaz Afzal1*, Muhammad Irfan Ullah2, Nadeem Ali3, Moeen Afzal1, Riaz Hussain4, Nabil A Alhakamy5, Nisreen Rajeh6, Sarmad Rehan7, Rehana Iqbal8, Muhammad Shahid Iqbal1, Ahrar Khan9*
1Department of Zoology, The Islamia University of Bahawalpur 63100, Pakistan
2Department of Pathobiology, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan
3Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah 21589, Saudi Arabia
4Department of Pathology, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur 63100, Pakistan
5Pharmaceutics Department, Faculty of Pharmacy, King Abdulaziz University, Jeddah 21589, Saudi Arabia
6Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah 21589, Saudi Arabia
7Department of Anatomy, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan
8Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan, Pakistan
9Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan

Abstract
Nanoparticles are used extensively in various industries, such as agriculture, food packaging, medical diagnostics and electronics. However, their increasing usage raises concerns regarding potential health hazards and environmental risks. This study examined the impact of intra-peritoneal injections of magnesium oxide (MgO) nanoparticles on the brain, testis, and muscles of male albino rats. Mature male rats (n=20) after acclimatization were randomly divided into four groups (G0, G1, G2, G3). The rats in the treated groups (G1-G3) were given MgO NPs @ 25 mg/kg, 50 mg/kg and 75 mg/kg respectively for ten consecutive days. G0 rats served as untreated control group. Results indicated that MgO NPs induced clinical alterations in exposed rats. The exposed organs including brain, and testis gained more weight and their stress parameters [reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS)] increased significantly in a dose dependent manner. Antioxidant enzymes including catalase (CAT), peroxidase (POD), reduced glutathione (GSH) and superoxide dismutase (SOD) reduced significantly in studied organs as compared to control ones. The treated rats have shown atrophy of neurons, microgliosis, cytoplasmic vacuolization, and congestion. Changes in the testis include inflammation, sloughing of cells, damaged spermatogonia, necrosis of spermatids, spermatogonia and arrest of spermatogenesis process. Conclusively, it is suggested that persistent application of nanomaterials at environmentally relevant concentrations may induce adverse toxicological effects in targeted and non-targeted exposed animals.

Keywords: Albino rats, MgO NPs, Oxidative stress, Antioxidant enzymes, Histopathology

Yasir Mahmood1, Nabeel Ijaz2, Aliza Maheen1, Ghulam Mustafa1, Duaa Abdullah Bafail3, Muhammad Rafi Qamar4, Muhammad Aitazaz Ahsan5, Nasir Masood6, Nisreen Rajeh7, Mudassar Mohiuddin8*
1Department of Zoology, The Islamia University, Bahawalpur, Pakistan
2Department of Clinical Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan
3Department of Clinical Pharmacology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
4Department of Clinical Medicine and Surgery, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan
5Department of Pathology, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan
6Department of Biosciences, COMSATS University Islamabad, Islamabad Campus, Pakistan
7Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
8Department of Microbiology, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan

Abstract
ZnO Nanoparticles (ZnO NPs) have wide applications in many fields of life ranging from health, food, agriculture, veterinary medicine, biotechnology, public health, textile and cosmetics. However, exposure to these NPs poses risks to public health, non-target living organisms and the environment. Hence, this study assessed toxicological impacts of ZnO NPs on hematopoietic tissues (bone marrow) and different visceral organs like lungs, intestine and muscles of male Wistar albino rats. Twenty male (20) Wistar albino rats were placed in four groups such as T0 (control group), T1 (50 mg/kg/day ZnO NPs), T2 (75 mg/kg/day ZnO NPs), and T3 (100 mg/kg ZnO NPs). Treated rats exhibited different signs of toxicity like depression and anxiety at higher doses of ZnO NPs. The bone marrow and other visceral organs/tissues were removed and analyzed to know the status of oxidative stress and antioxidant biomarkers. Results revealed notable increase (P≤0.05) in contents of oxidative stress biomarkers (ROS and TBARS) and significant decrease (P≤0.05) in antioxidant enzymes (POD, SOD, CAT, and GSH) in bone marrow as well as lungs, intestine, and muscles (gums) tissues. Histopathological examination indicated degeneration of muscle fibers, atrophied cells and presence of inflammatory materials in muscles (gums) of treated rats. Histologically, lungs were edematous, hemorrhagic and showed severe interstitial pneumonia while necrosis of epithelium of villi along with degeneration of villi in intestine of rats were observed at higher doses of nanoparticles. In conclusion, it can be suggested that ZnO-NPs may induce oxidative stress in multiple visceral organs of albino rats at higher concentrations highlighting disruption of physiological mechanisms.

Keywords: Nanoparticles, Zinc oxide, Nanoparticle toxicity, Oxidative stress biomarkers

Hira Ahsan^1,2, Maria Ayub¹, Mehraj Gul³, Amber Qureshi¹, Hani Z. Asfour⁴, Hafiz Muhammad Bilal⁵, Muhammad Azeem¹, Ammara Wahid¹, Nadeem Ali⁶, Rasheeha Naveed⁷, Mudasar Shabir⁸, Nisreen Rajeh⁹, Ayaz Mammadov¹⁰, Abu Baker Siddique^1*

¹Institute of Microbiology, Government College University, Faisalabad, Pakistan

²Academy of Medical Sciences, College of Henan Medicine, Zhengzhou University, Zhengzhou, Henan, 450052, China

³Quality Control Laboratory, National Institute of Health, Islamabad, Pakistan

⁴Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia

⁵Department of Rehabilitation and Allied Health sciences, Riphah International University, Lahore Campus, Lahore, Pakistan

⁶Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah-21589, Saudi Arabia

⁷Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan

⁸Institute of Advanced Materials and Flexible Electronics (IAMFE), Nanjing University of Information Science and Technology, Nanjing, China

⁹Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia

¹⁰Department of Life Sciences, Western Caspian University, Baku, Azerbaijan

Abstract

Multi-drug resistant (MDR) bacterial infections rapidly increase morbidity, mortality, and treatment options. Therefore, the search for, development of, or discovery of antimicrobial drugs capable of combating MDR bacteria is urgently needed. The potential of nanotechnology to advance nanomedicine for human health is being studied. The purpose of the present research is to investigate the antimicrobial activity of silver oxide nanoparticles against carbapenem-resistant Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. For this purpose, a total of 240 pus and wound samples were collected from the burn patients and further processed for isolation and identification of P. aeruginosa and MRSA according to standard microbiological techniques. Using a species-specific primer for each bacterial strain, polymerase chain reaction was used for molecular detection. Antimicrobial susceptibility was performed according to the Kirby-Bauer Disk Diffusion method. Molecular detection of carbapenemase-producing P. aeruginosa was performed by PCR by using specific primers. The agar well diffusion assay was used to examine the antibacterial properties of silver oxide nanoparticles, and the broth dilution assay was used to estimate the minimum inhibitory concentration and bactericidal concentration respectively. Out of 240 samples, 42 (17%) were identified as P. aeruginosa and 32 were confirmed as S. aureus isolates. From positive isolates of P. aeruginosa, 25 (59%) were recorded MDR P. aeruginosa and from positive isolates of S. aureus, 18 (56.25%) were detected as MRSA. The highly resistant drug against S. aureus was Penicillin G (100%) followed by Gentamicin (84.37%) and Ciprofloxacin (81.25%). The highly resistant drug against P. aeruginosa was Meropenem (100%), Imipenem (100%) followed by piperacillin (71.42%), gentamicin (64.28%), and ciprofloxacin (64.28%). Out of 42 P. aeruginosa isolates, 8(19%) the prevalence of carbapenemase encoding was noted as blaOXA 3(37.5%), blaNDM 2(25%), blaVIM 1(12.5%) blaKPC 1(12.5%) and blaIMP 1(12.5%).  Silver oxide nanoparticles were considered an effective antibacterial agent with 0.0065mg/mL-0.026mg/mL concentrations that highly inhibited the growth of MRSA and 0.39mg/mL-1.56mg/mL concentrations inhibited the growth of P. aeruginosa. The statistical analysis showed that the MIC and MBC for MDR P. aeruginosa were 0.96±0.43 μg/mL and 1.99±0.90 μg/mL, respectively, while for MRSA they were 0.01±0.008 μg/mL and 0.04±0.012 μg/mL, respectively. The MBC values were higher than MIC values for both pathogens. Silver oxide nanoparticles have such effective antibacterial properties that they can be used as an adequate source of antibacterial agents as alternatives to antibiotics.

Keywords: Silver oxide nanoparticles, Methicillin resistant, Gentamicin, Carbapenemase, Antimicrobial activity

Jingyu Feng^1,2, Li Yang¹, Jiguo Wang^1,2, Jing Zhang^1,2, Lizhu Lin^2,3*

¹Department of Oncology, Shenzhen Bao’an Traditional Chinese Medicine Hospital, Shenzhen 518133, China

²The First School of Clinical Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong, China

³Department of Oncology, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong, China

Abstract

The objective of this study was to elucidate the mechanism of the Hexuetongbi formula in treating Chemotherapy-induced Peripheral Neuropathy (CIPN) using network pharmacology, pharmacodynamics, and mechanistic approaches in an oxaliplatin-induced peripheral neuropathy rat model. Network pharmacology is crucial for understanding the multi-target effects of the Hexuetongbi formula on CIPN. This approach allows for a comprehensive mapping of the complex interactions between the formula’s constituents and the biological pathways involved in CIPN, revealing potential synergistic effects and enhancing the formula’s pharmacological validation. The Hexue Tongbi formula’s impact was analyzed on rats undergoing peripheral neuropathy, observing changes in the morphology of L4-6 dorsal root ganglion neurons through hematoxylin and eosin (HE) staining. Simultaneously, a network pharmacology approach was employed, utilizing TCMSP and GeneCards databases to identify common targets between CIPN and Hexue Tongbi’s therapeutic entities. These core targets were scrutinized through GO enrichment and KEGG pathway analysis. To validate the findings, mRNA and protein expression levels in the L4-6 dorsal root ganglion were examined using quantitative PCR and Western Blot assays. Significant modifications were observed in the frequency of cold stimulus withdrawal reflexes and the L4-6 dorsal root ganglion neurons, while the mechanical withdrawal reflex threshold displayed a considerable decrease. In rats treated with the Hexuetongbi formula post-oxaliplatin, there was noteworthy mitigation in the cold stimulation paw withdrawal threshold and augmentation in the mechanical stimulation paw withdrawal threshold. Network pharmacology identified 19 active constituents in Hexuetongbi and 35 targets for CIPN, with the PI3K/Akt signaling pathway emerging as a prominent target. There was a significant upregulation in PI3K, Akt1, Akt2, and Bcl-2 compared to controls, suggesting that Hexuetongbi effectively mitigates oxaliplatin-induced peripheral neuropathy through the modulation of the PI3K/Akt and Bcl-2 pathways.

Keywords: Oxaliplatin, Chemotherapy-induced peripheral neuropathy, Traditional Chinese medicine, Dorsal root ganglion, Apoptosis

Xinyou Liu^2,3, Yihong Luo¹, Zhenglin Wang¹, Cong Wang^1,2,3*

¹Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

²Department of General Surgery, Zhongshan Hospital (Xiamen), Fudan University, Xiamen 361015, China

³Xiamen Clinical Research Center for Cancer Therapy, Xiamen 361015, China

Abstract

This study addresses a critical knowledge gap in understanding the tumor microenvironment of thyroid cancer by elucidating the mechanism by which tumor cell-derived CD133-positive exosomes promote lymph node metastasis. We employed molecular docking, western blot, and other molecular characterizations to investigate the crucial interaction between CD133 and VEGFR3 and its impact on metastasis. Additionally, experiments assessed the influence of CD133-positive exosomes on lymphatic endothelial cell proliferation and migration.

Our findings demonstrate a significant direct interaction between CD133 and VEGFR3, as suggested by molecular docking. Furthermore, inhibition of CD133 expression resulted in a notable reduction in lymph node metastasis. We also observed that CD133-positive exosomes derived from thyroid cancer cells actively contribute to the migration and proliferation of lymphatic endothelial cells.

These results unveil a novel pathway for lymph node metastasis in thyroid cancer. The identification of CD133 and its interaction with VEGFR3 as key players in this process holds significant promise for the development of targeted therapeutic strategies.  By focusing on these targets, researchers can potentially improve the prognosis of patients with thyroid cancer.

Keywords: Thyroid cancer cells, Exomes, Lymph node metastasis, CD133, VEGFR3

Edith L. Kadege^1*, Pavithravani B. Venkataramana¹, Teshale Assefa², Joseph C. Ndunguru³, Jean Claude Rubyogo⁴, Ernest R. Mbega¹

¹School of Life Sciences and Bioengineering, The Nelson Mandela African Institution of Science and Technology, Arusha 447, Tanzania

²Crops for Nutrition and Health, International Center for Tropical Agriculture (CIAT), 2704 Arusha, Tanzania

³Department of Research and Innovation, Tanzania Agricultural Research Institute (TARI), Dodoma 1571, Tanzania
⁴Crops for Nutrition and Health, International Center for Tropical Agriculture (CIAT), Nairobi 823-00621, Kenya

Abstract

Tanzania is the top bean producer in Africa and seventh globally, exporting half of its beans to neighboring nations. This study determines common bean varietal performance in northern Tanzania. Disease infection and yield components of 22 genotypes were evaluated in on-station and on-farm trials. The study used a completely randomized factorial trial design with three replications to explore the individual and combined effects of genotype and environment on disease infection and grain yield in two on-station and six on-farm environments. Data were collected on number of emerging plants; canopy height, canopy width, plant vigor, disease infection levels, plant stands at harvest, number of pods per plant, number of grains per pod, 100-grain weight and grain yield and analyzed using R software. The combined analysis of variance revealed significant differences among genotypes, environment, and genotype by environment interactions. Bean canopy height, canopy width, plant vigor and grain yield were high at on-station trials, compared with on-farm trials. Advanced breeding lines showed 56% higher grain yield than commercial checks across study locations. Additive main effects and multiplicative interaction (AMMI) analysis revealed that, genotype was the dominant factor affecting common bean grain yields at 50.3%, whereas the environmental impacts were 25.7%. NUA 48 and NUA 64 were ideal genotypes showing anthracnose resistance and delivering higher grain yield. VTT 923-23-10 and Sweet Violet varieties were stable across the mega-environment. Therefore, NUA 48, NUA 64, VTT 923-23-10 and Sweet Violet are proposed for further evaluation within Tanzanian bean agroecosystem to identify farmers’ preferred varieties.

Keywords: Disease resistance, Yield components, Breeding lines, Bean, Tanzania

Sara Bano¹, Muhammad Imtiaz Rashid², Amna Akhtar¹, Farhan Hafeez¹, Rashid Nazir¹, Faridullah¹, Muhammad Irshad¹, Gabrijel Ondrasek^3*, Akhtar Iqbal^1*

¹Department of Environmental Sciences, COMSATS University Islamabad (CUI), Abbottabad Campus, Tobe Camp, University Road, Abbottabad, Pakistan.

²Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah 21589, Saudi Arabia.

³Department of Soil Amelioration, Faculty of Agriculture, University of Zagreb, HR-10000 Zagreb, Croatia.

Abstract

To enhance soil fertility and subsequent crop yields, digestate, byproduct of anaerobic digestion, can serve as a supplement or potential alternative to chemical fertilizers when sensibly utilized. This study assessed the impact of two types of digestates on two distinct soils for the perspective of agriculture (affecting pH, EC, organic carbon and mineral nitrogen) and environment (mineral nitrogen leaching under two rainfall patterns). Both soils mainly differed in silt and sand contents: 32% silt and 47% sand for soil-1 and 42% silt and 39% sand for soil-2. Two sets of controlled experiments served the purpose, in which first set involved a soil incubation experiment, applying two digestates to two soil types at 28°C for 60 days. The second set comprised reconstituted soil columns to collect soil solutions at depths of 2.5 cm and 7.5 cm after rainfall application under two patterns for 45 days. Results indicated that application of both digestates in test soils initially increased pH at day 15, followed by a decrease at days 30 and 60. Both digestates significantly elevated soil electrical conductivity compared to control treatments in both soils. Organic carbon content displayed variable impacts, with a slight decrease for solid digestate (12%) and higher decrease for liquid-amended soil (43%) for soil-1. While a significant decrease was observed for soil-2 throughout the incubation period for both amendments (34% and 36% for solid and liquid amended soils respectively). Rapid nitrification occurred with the application of both digestates in both soils, albeit at different rates. Soil-2 exhibited 1.2 to 2 folds higher net nitrification rate (depending upon digestate type and days of incubation) compared to soil-1. Liquid digestate induced more mineral nitrogen compared to solid digestate in both soils. Interestingly, rainfall frequency, digestate type, and soil type influenced the leaching of ammonium and nitrates, with nitrates recording higher levels in both soils, at both depths, and under both rainfall patterns.

 Keywords: Slurry, Nitrates, Crop productivity, Nitrification rate, Soil nutrition

Runglawan Sudmoon¹, Arunrat Chaveerach², Unchaleeporn Ameamsri², Penkhae Thamsenanupap^3,4, Natapol Pumipuntu^4,5, Shiou Yih Lee⁶, Ong Ghim Hock⁶, Tawatchai Tanee^3,4*

¹Faculty of Law, Khon Kaen University, Khon Kaen, Thailand

²Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand

³Faculty of Environment and Resource Studies, Mahasarakham University, Maha Sarakham, Thailand

⁴One Health Research Unit, Mahasarakham University, Maha Sarakham, Thailand

⁵Faculty of Veterinary Sciences, Mahasarakham University, Maha Sarakham, Thailand

⁶Faculty of Health and Life Sciences, INTI International University, Negeri Sembilan, Malaysia

Abstract

Heavy metal contamination, notably lead (Pb), poses a threat to plants and, consequently, human health through the food chain. This study investigates Pb accumulation in rice and how it affects the rice DNA. The rice (Oryza sativa) samples were cultivated in the soil supplemented with Pb at concentrations of 0, 15, 30, 60, and 120 mg/kg. Then the samples were harvested and analyzed for Pb accumulation and DNA alterations using the PCR amplification profiles measured through genomic template stability (GTS). The results demonstrate a Pb concentration hierarchy (root > stem > leaves), with Bioconcentration Factor (BCF) and Translocation Factor (TF) rising when soil Pb contents are supplemented. For DNA alterations, observed GTS values ranged from 23.3 to 76.67%, revealed a general decline with increasing Pb. This correlation assured the influence of Pb on rice DNA stability. Our findings suggest that heavy metal concentration, particularly Pb, has a direct influence on the integrity of rice DNA. Understanding these dynamics is vital for unraveling the complexities of heavy metal-induced genetic changes in plants and their potential implications for food safety and environmental health.

Keywords: Bioconcentration Factor, DNA changes, Genomic template stability, Lead accumulation, Translocation Factor

Rabia Kalsoom1, Hani Z. Asfour2, Hafiz Muhammad Ali3, Abdul Qayyum3*, Shazia Anjum4**, Faisal Maqbool1, Nuzhat Sial1, Riaz Hussain3, Sultan H. Alamri5, Nadeem Ali6, Nisreen Rajeh7, Irfan Irshad8, Asif Idrees9
1Department of Zoology, The Islamia University of Bahawalpur, 63100, Pakistan
2Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia
3Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, 63100, Pakistan
4Faculty of Chemical and Biological Sciences, The Islamia University of Bahawalpur, 63100, Pakistan
5Department of Family Medicine, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia
6Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah-21589, Saudi Arabia
7Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia
8Institute of Continuing Education and Extension, University of Veterinary and Animal Sciences, Lahore-54000, Pakistan
9KBCMA, College of Veterinary and Animal Sciences, Narowal, Pakistan

Abstract

Currently terrestrial ecologies are polluted by numerous chemical compounds unceasingly leading to high risk of exposure to variety of life. Among these, bifenthrin is widely used for the control of different sucking and chewing insects and other leaf miner insects around the globe. The current study elaborated the toxic effects of oral administration of bifenthrin in male Japanese quails at sub-lethal (less than 1% mortality) concentrations (10mg/kg, 20mg/kg and 30mg/kg b.w.) during a period of 30 days. During the course of the study, the quails did not show any behavioural or clinical signs. However, a significantly (p<0.05) decreased RBCs and platelets counts and haemoglobin concentration while a significantly (p<0.05) increased MCV, MCHC, total leucocytes and neutrophils were observed in the birds administered with higher concentrations of bifenthrin. Moreover, the incidence of lobed nuclei, blebbed nuclei, condensed nuclei, notched nuclei, binuclear, pear shaped and micro-nuclei were significantly (p<0.05) increased in the erythrocytes of the groups C and D during the experiment. The diameter of seminiferous tubules, height of germinal epithelium and the number of seminiferous tubules containing normal spermatozoa were significantly (p<0.01) decreased while the number of pyknotic cells and degenerated seminiferous tubules were increased significantly (p<0.05) towards the end of the experiment (day-30) in the quails of groups C and D compared to the control group. Hence, the alterations in the hematological indices and histopathological changes in heart, spleen and testes indicate potential toxicity of bifenthrin and its adverse effects in the Japanese quail even at sub-lethal concentrations.

 Keywords: Bifenthrin, Blood profile, Nuclear alterations, Seminiferous tubules, Histo-pathology, Testes

Mahmood Rasool^1*, Khalid I Alhassan², Sajjad Karim¹, Absarul Haque³, Mohammed HZ Mutwakil², Mohammed Alharthi⁴, Adeel G Chaudhary¹, Peter Natesan Pushparaj¹

¹Center of Excellence in Genomic Medicine Research, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia

²Department of Biological Science, King Abdulaziz University, Jeddah, Saudi Arabia

³King Fahd Medical Research Center, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia

⁴Faculty of Medicine, King Abdulaziz University Hospital, King Abdulaziz University, Jeddah, Saudi Arabia

Abstract

Despite considerable advancements in research, particularly in oncology, colorectal cancer (CRC) remains a formidable and deadly disease. It is crucial to delve deeper into the effects of targeted therapies, signaling pathways, and genetic regulation to ensure the effectiveness of cancer treatments. In this study, we obtained microarray data for four patient-derived CRC organoids from the Gene Expression Omnibus (GEO) database (accession number: GSE114060). We then used several next-generation knowledge discovery (NGKD) tools, such as GEO2R, Metascape, WebGestalt, and Ingenuity Pathway Analysis (IPA) software, to investigate the underlying molecular mechanisms in colorectal cancer-derived organoids treated with trametinib compared to those treated with DMSO. Our NGKD analysis revealed upregulation of SAMD, TP53, and SPTLC3 and downregulation of SPRED1, TGFBI, and DUSP6. The MAPK/ERK signaling pathway was significantly downregulated and was associated with reduced expression of CDX2 in CRC organoids treated with trametinib. We concluded that SAMD9, SPRED1, TGFBI, TP53, and DUSP6 are differentially regulated and can play a pivotal role in the downregulation of the MAPK/ERK signaling pathway to suppress CRC. The use of targeted therapies to regulate the specific gene signatures identified in the current study may be beneficial in the CRC associated tumor suppression . On the other hand, upregulation of SPTLC3 may be induced by MEK inhibitors and may cause hepatotoxicity alongside nonalcoholic steatohepatitis.

Keywords: Colorectal cancer, Organoids, Tumor suppression, MAPK/ERK signaling pathway, MEK inhibitor, Hepatotoxicity