Prompt engineering-enabled GenAI tools to map the landscape of crosstalk within the gut–brain axis (GBA) and its associated factors

Presently, the world is entangled by tools and technologies of artificial intelligence (AI). Everyone is now using AI tools and technologies. It is transforming the landscape of medicine and healthcare. In this context, Agha et al conveyed transparency standards for AI reporting in a critical article. They have provided detailed benchmarks, known as the TITAN guideline. It recommends adherence to these standards when employing AI in research[1]. The connection between the gut and the brain is very robust. The communication between these two components, known as the gut–brain axis (GBA), is well documented[2]. We can describe the relationship like an intense friendship. The communication between them influences the mental state and mood. The mental state and mood are also affected by the depth and extent of any disruption in the gut–brain relationship. They communicate and exchange signals, like two friends talking (crosstalk). A disruption in their friendship can lead to a bad mood[3,4]. Some research has included the gut microbiota in this axis. It is referred to as the brain–gut-microbiota axis[5,6]. However, it may seem surprising that when a person’s gut or stomach is healthy, their brain remains calm, leading to a better day. The gut is often called the body’s “second brain.” The gastrointestinal system is influenced by everything consumed daily. Therefore, the content of the food associated with it, such as its nutritional, fibrous, and other contents. Consuming unhealthy, non-nutritious foods puts additional strain on the intestines. It leads to deterioration of gut health, and the message is rapidly transmitted to the brain. When communication issues occur, they lead to a decline in mood and disrupt all activities. Scientists are investigating the intensity of the connection between the gut and the brain. It is believed that the stomach can influence emotions. However, there is a gut’s “network” with the gut hosted bacteria, a large community of beneficial and harmful bacteria. It is known as “gut microbiota.” Along with digestion, these communities produce metabolites associated with the GBA, which help maintain its balance. When this balance is disrupted, intestinal issues occur. Consequently, we feel the symptoms like stomach discomfort, fatigue, depression, mood fluctuations, and difficulty concentrating may arise. In the gastrointestinal tract, a diverse community of gut microbiota is found. It comprises a vast range of microbes, including both beneficial and harmful ones. It significantly influences the central nervous system via the GBA. This axis functions as a two-way communication channel between the gut microbiota and brain, enabling the microbiota to affect emotional and cognitive functions[4]. However, it is currently considered a crucial highway in the brain-gut-microbiota axis, which carries signals from gut microbes to the gut and, ultimately, to the brain (microbes-GBA)[7]. The vagus nerve, CN X, is a very complex nerve and the body’s longest nerve. Scientists found that the vagus nerve plays a vital role for the brain and gut communication. The nerve originates in the brain and reaches the intestines and other organs. Most signals from the gut are transmitted to the brain via the vagus nerve. Therefore, the nerve provides the brain with information about hunger, digestion, and the gut microbiota. Conversely, it was reported that about 20 percent of brain signals travel to the gut, influencing digestive processes[8–10]. Longo et al. illustrated that it plays a role in regulating T2DM (Type 2 diabetes mellitus) and obesity through its central regulation of metabolism and appetite secretion[11]. A neurotransmitter essential for brain regulation, and one crucial neurotransmitter is Serotonin. It is intricately connected to the gut. Moreover, the gut is more closely connected to the immune system through gut-microbiome–immune crosstalk[12]. Consequently, any disturbance any robust disturbance in gut might trigger inflammation. Subsequently, inflammatory agents can enter the bloodstream and reach the brain. It might potentially alter nerve function and affect the mental state. The microbiome-GBA is complex, with several other components and factors. The associated components and factors include gut microbiomes and microbial metabolites, neurotransmitters (involved in mood regulation) such as GABA (linked to stress management), the immune system, and neural connection pathways through the vagus nerve, etc. Among recent AI technologies, LLMs are a significant accomplishment. Following the release of ChatGPT, an LLM, it has become more widespread among professionals across different domains, from researchers to students[13,14]. Like LLMs, GenAI models can generate images, audio, videos, and other visual content. At the same time, to develop diverse models and provide solutions, researchers are using generative transformers. GenAI tools can be used for disease forecasting[15], drug discovery[16], and single-cell analysis across different diseases, such as Alzheimer’s[17]. Here, we have used prompt engineering-enabled GenAI tools, such as ChatGPT, Google Gemini, and Deepseek, to help understand the factors associated with landscape crosstalk within the GBA (Table 1). Scientists can use GenAI tools and technologies to gain deeper analysis into how these factors are influencing. GenAI tools and technologies can help map the brain’s influence on the gut, and the gut’s influence on the brain in a very profound manner. GenAI can analyze large-scale data from the microbiome-GBA and generate hypotheses with proper explanations. It will help in further investigation to understand the crosstalk between the “two friends” and their associated factors. Therefore, GenAI models will help reshape the understanding of mood and feelings by controlling through the GBA. Table 1 - Prompt engineering-enabled GenAI tools (ChatGPT, Google Gemini, and Deepseek) to understand the factors associated with the landscape crosstalk within the gut–brain axis (GBA) Sl no. Prompt engineering enabled questions ChatGPT Google Gemini Deepseek 1 Name and rank the factors associated with the gut-brain axis, which influences mental state and mood. 1. Gut microbiota composition Tier 1: Core modulators and pathways Tier 1: Core physiological & neural pathways 2. Diet and nutrition 3. Inflammation and immune activation These factors are fundamental to the existence and function of the gut-brain axis and have the most far-reaching influence on mental health These are the essential biological communication highways that make the axis possible. 4. Vagus nerve activity 5. Microbial metabolites 6. Stress and HPA (hypothalamic–pituitary–adrenal) axis 1. Gut microbiota (dysbiosis) 1. The vagus nerve 7. Gut barrier integrity (“Leaky Gut”) 2. Vagus nerve 2. Neurotransmitter production 8. Neurotransmitters produced in the gut 3. Neurotransmitters and metabolites 3. Immune-mediated signaling (inflammation) 9. Hormones and peptides Tier 2: Systemic regulators and mediators Tier 2: Microbial & metabolic factors 10. Genetics and epigenetics These physiological systems are critical for relaying and executing the signals passed between the gut and brain, especially in response to stress and inflammation These are the dynamic inhabitants and their chemical outputs that drive much of the modulation 11. Antibiotics and medication use 12. Sleep and circadian rhythms 13. Physical activity 4. Microbiota composition & diversity 5. Microbial metabolites 14. Early-life microbial exposure 4. Hypothalamic–pituitary–adrenal (HPA) axis 6. Tryptophan metabolism Tier 3: Structural & barrier factors 5. Immune system/inflammation The integrity of the gut itself determines signal quality 6. Enteric nervous system (ENS) 7. Intestinal permeability & barrier function Tier 3: Key environmental/lifestyle factors 8. Enteroendocrine cell signaling While not part of the biological axis itself, these external factors are vital because they directly and rapidly modify the Tier 1 and 2 components, leading to changes in mental state Tier 4: Modulating & lifestyle factors These are external inputs that shape the above tiers. 9. Diet 10. Stress (acute & chronic) 7. Diet/nutrition 11. Medications (especially antibiotics) 8. Stress and sleep 12. Sleep & circadian rhythms 13. Exercise 2 List the top five factors the gut–brain axis. 1. Gut microbiota composition 1. Gut microbiota and dysbiosis 1. The vagus nerve 2. Diet and nutrition 2. Vagus nerve signaling 2. Gut microbiota composition & diversity 3. Vagus nerve signaling 3. Neurotransmitters and microbial metabolites 3. Systemic inflammation 4. Inflammation and immune activation 4. Microbial metabolites (short-chain fatty acids – SCFAs) 5. Stress and the HPA axis 4. HPA axis regulation 5. Immune activation and inflammation 5. Intestinal barrier integrity (“Leaky Gut”)