The project, which will run until the end of March next year, will see the charity work across eight areas of England and Wales to identify, support and protect male victims of sexual abuse and exploitation.
The charity said the project is necessary due to a lack of specialist support in many areas for boys who are victims of abuse and exploitation.
The Youth Justice Board, youth offending teams and male sexual exploitation programme Blast are also involved in developing the project.
Barnardo's estimates that a third of exploitation victims are male, adding that vulnerable boys who are prone to mental health issues, substance abuse, homelessness, social isolation and offending are particularly at risk.
"We welcome this funding, which will enable Barnardo's to develop a greater understanding of the assessment and support required for boys and young men with a background of sexual abuse and trauma," Barnardo's chief executive Javed Khan said.
"Barnardo's has identified that young male victims of sexual abuse are under-represented in services supporting abuse victims, whilst over-represented within the criminal justice system.
"An early experience of sexual abuse can lead boys to getting involved in risky behaviour and exploitation. This work will help vulnerable boys and young men get the support they need, when they need it."
The project will operate in the police and crime commissioner areas of: West Midlands, West Mercia, Warwickshire, Avon and Somerset, South Wales, North Wales, Dyfed and Gwent.
A report released by Barnardo's Cymru last year found that boys who are victims of sexual exploitation and abuse often miss out on seeking help as they are reluctant to talk about their experiences.
That report also said that more training among children's professionals was needed to better identify male victims.
The project is one of four to share £600,000 in Home Office funding handed out today to organisations that support victims of child sexual abuse.
The National Association of People Abused in Childhood (NAPAC) received £314,360, Safeline Warwick was handed £2,099 and Rape Crisis England and Wales gained £76,659.
"Child sexual abuse is a devastating crime and we are determined to ensure every victim has the support they need, as we work to bring their abusers to justice," Sarah Newton, minister for crime, safeguarding and vulnerability, said.
"This government is committed to protecting the most vulnerable in our society and we have done more than any other to tackle these horrific offences.
"We have increased support for victims of sexual abuse, invested in training and technology to improve law enforcement's response to abuse both on and offline, and brought in a tougher inspection regime to ensure all frontline professions are meeting their child protection duties.
"But there is more to do, and this funding will ensure that charities who protect and support people who have suffered child sexual abuse are able to continue and develop the vital support they offer."
