Low back pain (LBP) is a common condition that can affect athletes of all ages. The risk factors for LBP onset and worsening associated with the lacrosse shooting motion are not yet known. To identify training and biomechanical factors associated with preexisting LBP and development of LBP over 6 months in youth, high school, and collegiate lacrosse players. Case-control study; Level of evidence, 3. A total of 128 lacrosse players were enrolled in this study between January 2016 and January 2019. Player characteristics, lacrosse experience, and participation in other sports were self-reported. At baseline and 2-, 4-, and 6-month follow-ups, the players self-rated the presence and severity of LBP using a numeric pain rating scale (0-10 points). Participants were grouped according to LBP symptoms: no LBP at any time point (n = 102), preexisting LBP (n = 17), or developed LBP within the 6-month period (n = 9). The lacrosse shooting motion was captured via 3-dimensional motion analysis, and kinematic and kinetic variables were recorded. A Low Back Stress Index was used to estimate lumbar stress as a function of pelvic acceleration at the time of maximum lateral trunk lean during the shot. Univariate analyses of covariance and logistic regression models were used to address study aims. Compared with the no-LBP group, the preexisting LBP group demonstrated 13.9% to 22.9% lower maximum angular velocities at the pelvis, trunk, and shoulders in the transverse plane (P < .05), 19.3% less collective pelvis-shoulder rotation in the transverse plane (P = .015), and 4.5% more knee flexion excursion (P = .063). The developed-LBP group produced 2.3% to 11.1% higher angular velocities in the pelvis, trunk, and shoulder and generated maximum pelvic acceleration values 36% to 42% higher than the remaining groups (P < .05 for both). Mean Low Back Stress Index values were not statistically significant among the groups (no LBP: 12,504 ± 13,076 deg2/s2; preexisting LBP: 8808 ± 10,174 deg2/s2; developed LBP: 19,389 ± 13,590 deg2/s2; P = .157). Preexisting LBP was associated with significantly restricted motion of the pelvis, trunk, and shoulders during a lacrosse shot. Excessive pelvic acceleration may be related to the development of LBP in lacrosse players.