gh-k-dense-ai-claude-scientific-skills-scientific-skills/skills/scvi-tools/references/theoretical-foundations.md

Theoretical Foundations of scvi-tools

This document explains the mathematical and statistical principles underlying scvi-tools.

Core Concepts

Variational Inference

What is it? Variational inference is a technique for approximating complex probability distributions. In single-cell analysis, we want to understand the posterior distribution p(z|x) - the probability of latent variables z given observed data x.

Why use it?

• Exact inference is computationally intractable for complex models
• Scales to large datasets (millions of cells)
• Provides uncertainty quantification
• Enables Bayesian reasoning about cell states

How does it work?

1. Define a simpler approximate distribution q(z|x) with learnable parameters
2. Minimize the KL divergence between q(z|x) and true posterior p(z|x)
3. Equivalent to maximizing the Evidence Lower Bound (ELBO)

ELBO Objective:

ELBO = E_q[log p(x|z)] - KL(q(z|x) || p(z))
       ↑                    ↑
  Reconstruction          Regularization

• Reconstruction term: Model should generate data similar to observed
• Regularization term: Latent representation should match prior

Variational Autoencoders (VAEs)

Architecture:

x (observed data)
    ↓
[Encoder Neural Network]
    ↓
z (latent representation)
    ↓
[Decoder Neural Network]
    ↓
x̂ (reconstructed data)

Encoder: Maps cells (x) to latent space (z)

• Learns q(z|x), the approximate posterior
• Parameterized by neural network with learnable weights
• Outputs mean and variance of latent distribution

Decoder: Maps latent space (z) back to gene space

• Learns p(x|z), the likelihood
• Generates gene expression from latent representation
• Models count distributions (Negative Binomial, Zero-Inflated NB)

Reparameterization Trick:

• Allows backpropagation through stochastic sampling
• Sample z = μ + σ ⊙ ε, where ε ~ N(0,1)
• Enables end-to-end training with gradient descent

Amortized Inference

Concept: Share encoder parameters across all cells.

Traditional inference: Learn separate latent variables for each cell

• n_cells × n_latent parameters
• Doesn't scale to large datasets

Amortized inference: Learn single encoder for all cells

• Fixed number of parameters regardless of cell count
• Enables fast inference on new cells
• Transfers learned patterns across dataset

Benefits:

• Scalable to millions of cells
• Fast inference on query data
• Leverages shared structure across cells
• Enables few-shot learning

Statistical Modeling

Count Data Distributions

Single-cell data are counts (integer-valued), requiring appropriate distributions.

Negative Binomial (NB)

x ~ NB(μ, θ)

• μ (mean): Expected expression level
• θ (dispersion): Controls variance
• Variance: Var(x) = μ + μ²/θ

When to use: Gene expression without zero-inflation

• More flexible than Poisson (allows overdispersion)
• Models technical and biological variation

Zero-Inflated Negative Binomial (ZINB)

x ~ π·δ₀ + (1-π)·NB(μ, θ)

• π (dropout rate): Probability of technical zero
• δ₀: Point mass at zero
• NB(μ, θ): Expression when not dropped out

When to use: Sparse scRNA-seq data

• Models technical dropout separately from biological zeros
• Better fit for highly sparse data (e.g., 10x data)

Poisson

x ~ Poisson(μ)

• Simplest count distribution
• Mean equals variance: Var(x) = μ

When to use: Less common; ATAC-seq fragment counts

• More restrictive than NB
• Faster computation

Batch Correction Framework

Problem: Technical variation confounds biological signal

• Different sequencing runs, protocols, labs
• Must remove technical effects while preserving biology

scvi-tools approach:

1. Encode batch as categorical variable s
2. Include s in generative model
3. Latent space z is batch-invariant
4. Decoder conditions on s for batch-specific effects

Mathematical formulation:

Encoder: q(z|x, s)  - batch-aware encoding
Latent: z           - batch-corrected representation
Decoder: p(x|z, s)  - batch-specific decoding

Key insight: Batch info flows through decoder, not latent space

• z captures biological variation
• s explains technical variation
• Separable biology and batch effects

Deep Generative Modeling

Generative model: Learns p(x), the data distribution

Process:

1. Sample latent variable: z ~ p(z) = N(0, I)
2. Generate expression: x ~ p(x|z)
3. Joint distribution: p(x, z) = p(x|z)p(z)

Benefits:

• Generate synthetic cells
• Impute missing values
• Quantify uncertainty
• Perform counterfactual predictions

Inference network: Inverts generative process

• Given x, infer z
• q(z|x) approximates true posterior p(z|x)

Model Architecture Details

scVI Architecture

Input: Gene expression counts x ∈ ℕ^G (G genes)

Encoder:

h = ReLU(W₁·x + b₁)
μ_z = W₂·h + b₂
log σ²_z = W₃·h + b₃
z ~ N(μ_z, σ²_z)

Latent space: z ∈ ℝ^d (typically d=10-30)

Decoder:

h = ReLU(W₄·z + b₄)
μ = softmax(W₅·h + b₅) · library_size
θ = exp(W₆·h + b₆)
π = sigmoid(W₇·h + b₇)  # for ZINB
x ~ ZINB(μ, θ, π)

Loss function (ELBO):

L = E_q[log p(x|z)] - KL(q(z|x) || N(0,I))

Handling Covariates

Categorical covariates (batch, donor, etc.):

• One-hot encoded: s ∈ {0,1}^K
• Concatenate with latent: [z, s]
• Or use conditional layers

Continuous covariates (library size, percent_mito):

• Standardize to zero mean, unit variance
• Include in encoder and/or decoder

Covariate injection strategies:

• Concatenation: [z, s] fed to decoder
• Deep injection: s added at multiple layers
• Conditional batch norm: Batch-specific normalization

Advanced Theoretical Concepts

Transfer Learning (scArches)

Concept: Use pretrained model as initialization for new data

Process:

1. Train reference model on large dataset
2. Freeze encoder parameters
3. Fine-tune decoder on query data
4. Or fine-tune all with lower learning rate

Why it works:

• Encoder learns general cellular representations
• Decoder adapts to query-specific characteristics
• Prevents catastrophic forgetting

Applications:

• Query-to-reference mapping
• Few-shot learning for rare cell types
• Rapid analysis of new datasets

Multi-Resolution Modeling (MrVI)

Idea: Separate shared and sample-specific variation

Latent space decomposition:

z = z_shared + z_sample

• z_shared: Common across samples
• z_sample: Sample-specific effects

Hierarchical structure:

Sample level: ρ_s ~ N(0, I)
Cell level: z_i ~ N(ρ_{s(i)}, σ²)

Benefits:

• Disentangle biological sources of variation
• Compare samples at different resolutions
• Identify sample-specific cell states

Counterfactual Prediction

Goal: Predict outcome under different conditions

Example: "What would this cell look like if from different batch?"

Method:

1. Encode cell to latent: z = Encoder(x, s_original)
2. Decode with new condition: x_new = Decoder(z, s_new)
3. x_new is counterfactual prediction

Applications:

• Batch effect assessment
• Predicting treatment response
• In silico perturbation studies

Posterior Predictive Distribution

Definition: Distribution of new data given observed data

p(x_new | x_observed) = ∫ p(x_new|z) q(z|x_observed) dz

Estimation: Sample z from q(z|x), generate x_new from p(x_new|z)

Uses:

• Uncertainty quantification
• Robust predictions
• Outlier detection

Differential Expression Framework

Bayesian Approach

Traditional methods: Compare point estimates

• Wilcoxon, t-test, etc.
• Ignore uncertainty
• Require pseudocounts

scvi-tools approach: Compare distributions

• Sample from posterior: μ_A ~ p(μ|x_A), μ_B ~ p(μ|x_B)
• Compute log fold-change: LFC = log(μ_B) - log(μ_A)
• Posterior distribution of LFC quantifies uncertainty

Bayes Factor

Definition: Ratio of posterior odds to prior odds

BF = P(H₁|data) / P(H₀|data)
     ─────────────────────────
     P(H₁) / P(H₀)

Interpretation:

• BF > 3: Moderate evidence for H₁
• BF > 10: Strong evidence
• BF > 100: Decisive evidence

In scvi-tools: Used to rank genes by evidence for DE

False Discovery Proportion (FDP)

Goal: Control expected false discovery rate

Procedure:

1. For each gene, compute posterior probability of DE
2. Rank genes by evidence (Bayes factor)
3. Select top k genes such that E[FDP] ≤ α

Advantage over p-values:

• Fully Bayesian
• Natural for posterior inference
• No arbitrary thresholds

Implementation Details

Optimization

Optimizer: Adam (adaptive learning rates)

• Default lr = 0.001
• Momentum parameters: β₁=0.9, β₂=0.999

Training loop:

1. Sample mini-batch of cells
2. Compute ELBO loss
3. Backpropagate gradients
4. Update parameters with Adam
5. Repeat until convergence

Convergence criteria:

• ELBO plateaus on validation set
• Early stopping prevents overfitting
• Typically 200-500 epochs

Regularization

KL annealing: Gradually increase KL weight

• Prevents posterior collapse
• Starts at 0, increases to 1 over epochs

Dropout: Random neuron dropping during training

• Default: 0.1 dropout rate
• Prevents overfitting
• Improves generalization

Weight decay: L2 regularization on weights

• Prevents large weights
• Improves stability

Scalability

Mini-batch training:

• Process subset of cells per iteration
• Batch size: 64-256 cells
• Enables scaling to millions of cells

Stochastic optimization:

• Estimates ELBO on mini-batches
• Unbiased gradient estimates
• Converges to optimal solution

GPU acceleration:

• Neural networks naturally parallelize
• Order of magnitude speedup
• Essential for large datasets

Connections to Other Methods

vs. PCA

• PCA: Linear, deterministic
• scVI: Nonlinear, probabilistic
• Advantage: scVI captures complex structure, handles counts

vs. t-SNE/UMAP

• t-SNE/UMAP: Visualization-focused
• scVI: Full generative model
• Advantage: scVI enables downstream tasks (DE, imputation)

vs. Seurat Integration

• Seurat: Anchor-based alignment
• scVI: Probabilistic modeling
• Advantage: scVI provides uncertainty, works for multiple batches

vs. Harmony

• Harmony: PCA + batch correction
• scVI: VAE-based
• Advantage: scVI handles counts natively, more flexible

Mathematical Notation

Common symbols:

• x: Observed gene expression (counts)
• z: Latent representation
• θ: Model parameters
• q(z|x): Approximate posterior (encoder)
• p(x|z): Likelihood (decoder)
• p(z): Prior on latent variables
• μ, σ²: Mean and variance
• π: Dropout probability (ZINB)
• θ (in NB): Dispersion parameter
• s: Batch/covariate indicator

Further Reading

Key Papers:

1. Lopez et al. (2018): "Deep generative modeling for single-cell transcriptomics"
2. Xu et al. (2021): "Probabilistic harmonization and annotation of single-cell transcriptomics"
3. Boyeau et al. (2019): "Deep generative models for detecting differential expression in single cells"

Concepts to explore:

• Variational inference in machine learning
• Bayesian deep learning
• Information theory (KL divergence, mutual information)
• Generative models (GANs, normalizing flows, diffusion models)
• Probabilistic programming (Pyro, PyTorch)

Mathematical background:

• Probability theory and statistics
• Linear algebra and calculus
• Optimization theory
• Information theory