Angioimmunoblastic T-cell lymphoma (AITL) is a uniquely aggressive mature T-cell neoplasm. In recent years, recurrent genetic mutations in ras homolog family member A (
RHOA
), tet methylcytosine dioxygenase 2 (
TET2
), DNA methyltransferase 3 alpha (
DNMT3A
) and isocitrate dehydrogenase [NADP(+)] 2 (
IDH2
) have been identified as associated with AITL. However, a deep molecular study assessing both DNA mutations and RNA expression profile combined with digital image analysis is lacking. The present study aimed to evaluate the significance of molecular and morphologic features by high resolution digital image analysis in several cases of AITL. To do so, a total of 18 separate tissues from 10 patients with AITL were collected and analyzed. The results identified recurrent mutations in
RHOA
,
TET2
,
DNMT3A
, and
IDH2
, and demonstrated increased DNA mutations in coding, promoter and CCCTC binding factor (CTCF) binding sites in
RHOA
mutated AITLs vs.
RHOA
non-mutated cases, as well as increased overall survival in
RHOA
mutated patients. In addition, single cell computational digital image analysis morphologically characterized
RHOA
mutated AITL cells as distinct from cells from
RHOA
mutation negative patients. Computational analysis of single cell morphological parameters revealed that
RHOA
mutated cells have decreased eccentricity (more circular) compared with
RHOA
non-mutated AITL cells. In conclusion, the results from the present study expand our understanding of AITL and demonstrate that there are specific cell biological and morphological manifestations of
RHOA
mutations in cases of AITL.