The senators argue that Nvidia's practice of bundling software and hardware stifles innovation and locks in customers.
Demand Progress, along with nine other groups, has written to the Department of Justice's antitrust chief, Jonathan Kanter, calling for a probe into Nvidia's business practices.
Nvidia's market value soared to $3 trillion this summer, driven by demand for chips capable of running complex generative AI models.
The groups, which advocate against monopolies and for government oversight of tech companies, have criticised Nvidia's bundling practices, which have also drawn the attention of French antitrust authorities.
"This aggressively proprietary approach, which is strongly contrary to industry norms about collaboration and interoperability, acts to lock in customers and stifles innovation," the groups stated.
Nvidia controls approximately 80 per cent of the AI chip market, including custom AI processors made by cloud computing giants like Google, Microsoft, and Amazon. These chips are typically rented through each platform rather than sold directly.
A spokesperson for Nvidia responded, "Regulators need not be concerned, as we scrupulously adhere to all laws and ensure that Nvidia is openly available in every cloud and on-premises for every enterprise. We will continue to support aspiring innovators in every industry and market and are happy to provide any information regulators need."