Self-supervised speech representation learning has shown promising results in various speech processing tasks. However, the pre-trained models, e.g., HuBERT, are storage-intensive Transformers, limiting their scope of applications under lowresource settings. To this end, we propose LightHuBERT, a once-for-all Transformer compression framework, to find the desired architectures automatically by pruning structured parameters. More precisely, we create a Transformer-based supernet that is nested with thousands of weight-sharing subnets and design a two-stage distillation strategy to leverage the contextualized latent representations from HuBERT. Experiments on automatic speech recognition (ASR) and the SU-PERB benchmark show the proposed LightHuBERT enables over 10 9 architectures concerning the embedding dimension, attention dimension, head number, feed-forward network ratio, and network depth. LightHuBERT outperforms the original HuBERT on ASR and five SUPERB tasks with the Hu-BERT size, achieves comparable performance to the teacher model in most tasks with a reduction of 29% parameters, and obtains a 3.5× compression ratio in three SUPERB tasks, e.g., automatic speaker verification, keyword spotting, and intent classification, with a slight accuracy loss. The code and pre-trained models are available at https://github.com/ mechanicalsea/lighthubert.